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Why are there only 6 molecules of water produced in the synthesis of the fatty acid, palmitate?

Why are there only 6 molecules of water produced in the synthesis of the fatty acid, palmitate?


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The equation for biosynthesis is

1 Acetyl-CoA + 7 Malonyl-CoA + 14 NADPH + 14H+ -> Palmitate + 7 CO2 + 14 NADP+ + 8HS-CoA + 6 H2O

I really don't understand how there is only 6 H2O produced… and not 7?

Every cycle, there is the condensation of one molecule of water.

The first turn yields a 4 carbon molecules. Since palmitate is a 16C fatty acid, and two carbons are added each turn, there should be 6 turns remaining, for a total of 7 turn.

All the other numbers seem right. At first I thought my teacher could have made a mistake but everywhere I look on the net, it say 6 H2O.

Someone has an explanation for this?

Thank you :)

Edit: I found the answer in Voet & Voet yesterday. I add it here in case it can help someone else!

At the end of the biosynthesis, the palmitate is bonded with the Acyl-Carrier Protein (ACP) and it takes an hydrolysis to separate it, hence the missing H2O.


I found the answer in Voet & Voet yesterday.

At the end of the biosynthesis, the palmitate is bonded with the Acyl-Carrier Protein (ACP) and it takes an hydrolysis to separate it, hence the missing H2O.


2.7: Fatty Acids

  • Contributed by E. V. Wong
  • Axolotl Academica Publishing (Biology) at Axolotl Academica Publishing

Unlike monosaccharides, nucleotides, and amino acids, fatty acids are not monomers that are linked together to form much larger molecules. Although fatty acids can be linked together, for example, into triacylglycerols or phospholipids, they are not linked directly to one another, and generally no more than three in a given molecule. The fatty acids themselves are long chains of carbon atoms topped off with a carboxyl group. The length of the chain can vary, although most are between 14 and 20 carbons, and in higher order plants and animals, fatty acids with 16 and 18 carbons are the major species.

Figure (PageIndex<13>). Fatty acids. (Top) Stearic acid is a fully saturated fatty acid with no carbon-carbon double bonds. (Bottom) Oleic acid is an unsaturated fatty acid.

Due to the mechanism of synthesis, most fatty acids have an even number of carbons, although odd-numbered carbon chains can also be generated. More variety can be generated by double-bonds between the carbons. Fatty acid chains with no double bonds are saturated, because each carbon is saturated with as many bonded hydrogen atoms as possible. Fatty acid chains with double bonds are unsaturated (Figure (PageIndex<13>)). Those with more than one double bond are called polyunsaturated. The fatty acids in eukaryotic cells are nearly evenly divided between saturated and unsaturated types, and many of the latter may be polyunsaturated. In prokaryotes, polyunsaturation is rare, but other modifications such as branching and cyclization are more common than in eukaryotes. A table of common fatty acids is shown below.

Myristic Acid 14:0 (14 carbons, no double bonds
Palmitic Acid 16:0
Stearic Acid 18:0
Arachidic Acid 20:0
Palmitoleic Acid 16:1
Oleic Acid 18:1
Linoleic Acid 18:2
Arachidonic Acid 2:4

There are significant physical differences between the saturated and unsaturated fatty acids due simply to the geometry of the double-bonded carbons. A saturated fatty acid is very flexible with free rotation around all of its C-C bonds. The usual linear diagrams and formulas depicting saturated fatty acids also serve to explain the ability of saturated fatty acids to pack tightly together, with very little intervening space. Unsaturated fatty acids, on the other hand are unable to pack as tightly because of the rotational constraint impoarted by the double bond. The carbons cannot rotate around the double bond, so there is now a &ldquokink&rdquo in the chain. Generally, double-bonded carbons in fatty acids are in the cis- configuration, introducing a 30-degree bend in the structure.

Figure (PageIndex<14>). Triglycerides. These lipids are formed by conjugation of a glycerol to three fatty acyl chains through ester bonds from each glycerol oxygen.

Fatty acids inside cells are usually parts of larger molecules, rather than free acids. Some of the most common lipids derived from fatty acids are triacylglycerols, phosphoglycerides, and sphingolipids. Triacylglycerols, as the name implies, is three fatty acid (acyl) chains connected to a glycerol molecule by ester bonds (Figure (PageIndex<14>)). Triacylglycerols, also known as triglycerides, may have fatty acids of the same (simple triacylglycerols) or varying types (mixed triacylglycerols). Mixtures of these are the primary long-term energy storage molecules for most organisms. Although they may be referred to colloquially as fats or oils, the only real difference is the degree of saturation of their constituent fatty acids. Mixtures with higher percentages of saturated fatty acids have a higher melting point and if they are solid at room temperature, they are referred to as fats. Triacylglycerol mixtures remaining liquid at room temperature are oils.

In human medicine, a common test for heart disease risk factors is measurement of triglyceride levels in the blood. Although various cell types can make and use triglycerides, most of the triglycerides in people are concentrated in the adipose tissue, which is made up of adipocytes, or fat cells, though liver is also a significant fat store. These cells have specialized to carry fat globules that take up most of the volume of the cell. When triglyceride levels in the blood are high, it means that fat is being produced or ingested faster than it can be taken up by the adipocytes.

Figure (PageIndex<15>). A phospholipid: the glycerol backbone (red) connects to two fatty acids and to a phosphate and polar head group.

Phospholipids (also called phosphoglycerides or glycerophospholipids), are also based on attachment of fatty acids to glycerol. However, instead of three fatty acyl tails, there are only two, and in the third position is a phosphate group (Figure (PageIndex<15>)). The phosphate group also attaches to a &ldquohead group&rdquo . The identity of the head group names the molecule, along with the fatty acyl tails. In the example Figure, 1-stearoyl refers to the stearic acid on the 1-carbon of the glycerol backbone 2-palmitoyl refers to the palmitic acid on the 2-carbon of the glycerol, and phosphatidylethanolamine refers to the phosphate group and its attached ethanolamine, that are linked to the glycerol 3-carbon. Because of the negatively-charge phosphate group, and a head group that is often polar or charged, phospholipids are amphipathic - carrying a strong hydrophobic character in the two fatty acyl tails, and a strong hydrophilic character in the head group. This amphipathicity is crucial in the role of phospholipids as the primary component of cellular membranes.

Figure (PageIndex<16>). Sphingolipids are based on the amino alcohol, sphingosine (A). Ceramides have a fatty acid tail attached, and a ceramide with a phosphocholine head group is a sphingomyelin (B). If the head group is a sugar, then the molecule is a cerebroside. (C)

Sphingolipids (Figure (PageIndex<16>)) are also important constituents of membranes, and are based not upon a glycerol backbone, but on the amino alcohol, sphingosine (or dihydrosphingosine). There are four major types of sphingolipids: ceramides, sphingomyelins, cerebrosides, and gangliosides. Ceramides are sphingosine molecules with a fatty acid tail attached to the amino group. Sphingomyelins are ceramides in which a phosphocholine or phosphoethanolamine are attached to the 1-carbon. Cerebrosides and gangliosides are glycolipids - they have a sugar or sugars, respectively, attached to the 1-carbon of a ceramide. The oligosaccharides attached to gangliosides all contain at least one sialic acid residue. In additional to being a structural component of the cell membrane, gangliosides are particular important in cell to cell recognition.

Lipids are vaguely defined as biological compounds that are insoluble in water but are soluble in organic solvents such as methanol or chloroform. This includes the fatty acid derivatives listed above, and it includes the final topic for this chapter, cholesterol. Cholesterol (Figure (PageIndex<17>)) is the major biological derivative of cyclopentanoperhydrophenanthrene, a saturated hydrocarbon consisting of four fused ring formations. It is an important component of plasma membranes in animal cells, and is also the metabolic precursor to steroid hormones, such as cortisol or b-estradiol. Plant cells have little if any cholesterol, but other sterols like stigmasterol are present. Similarly, fungi have their particular sterols. However, prokaryotes do not, for the most part, contain any sterol molecules.


Contents

Straight-chain fatty acids occur in two types: saturated and unsaturated.

Saturated straight-chain fatty acids Edit

Much like β-oxidation, straight-chain fatty acid synthesis occurs via the six recurring reactions shown below, until the 16-carbon palmitic acid is produced. [1] [2]

The diagrams presented show how fatty acids are synthesized in microorganisms and list the enzymes found in Escherichia coli. [1] These reactions are performed by fatty acid synthase II (FASII), which in general contain multiple enzymes that act as one complex. FASII is present in prokaryotes, plants, fungi, and parasites, as well as in mitochondria. [3]

In animals, as well as some fungi such as yeast, these same reactions occur on fatty acid synthase I (FASI), a large dimeric protein that has all of the enzymatic activities required to create a fatty acid. FASI is less efficient than FASII however, it allows for the formation of more molecules, including "medium-chain" fatty acids via early chain termination. [3]

Once a 16:0 carbon fatty acid has been formed, it can undergo a number of modifications, resulting in desaturation and/or elongation. Elongation, starting with stearate (18:0), is performed mainly in the ER by several membrane-bound enzymes. The enzymatic steps involved in the elongation process are principally the same as those carried out by FAS, but the four principal successive steps of the elongation are performed by individual proteins, which may be physically associated. [4] [5]

Note that during fatty synthesis the reducing agent is NADPH, whereas NAD is the oxidizing agent in beta-oxidation (the breakdown of fatty acids to acetyl-CoA). This difference exemplifies a general principle that NADPH is consumed during biosynthetic reactions, whereas NADH is generated in energy-yielding reactions. [6] (Thus NADPH is also required for the synthesis of cholesterol from acetyl-CoA while NADH is generated during glycolysis.) The source of the NADPH is two-fold. When malate is oxidatively decarboxylated by "NADP + -linked malic enzyme" to form pyruvate, CO2 and NADPH are formed. NADPH is also formed by the pentose phosphate pathway which converts glucose into ribose, which can be used in synthesis of nucleotides and nucleic acids, or it can be catabolized to pyruvate. [6]

Conversion of carbohydrates into fatty acids Edit

In humans, fatty acids are formed from carbohydrates predominantly in the liver and adipose tissue, as well as in the mammary glands during lactation.

The pyruvate produced by glycolysis is an important intermediary in the conversion of carbohydrates into fatty acids and cholesterol. [6] This occurs via the conversion of pyruvate into acetyl-CoA in the mitochondrion. However, this acetyl CoA needs to be transported into cytosol where the synthesis of fatty acids and cholesterol occurs. This cannot occur directly. To obtain cytosolic acetyl-CoA, citrate (produced by the condensation of acetyl CoA with oxaloacetate) is removed from the citric acid cycle and carried across the inner mitochondrial membrane into the cytosol. [6] There it is cleaved by ATP citrate lyase into acetyl-CoA and oxaloacetate. The oxaloacetate can be used for gluconeogenesis (in the liver), or it can be returned into mitochondrion as malate. [7] The cytosolic acetyl-CoA is carboxylated by acetyl CoA carboxylase into malonyl CoA, the first committed step in the synthesis of fatty acids. [7] [8]

Animals cannot resynthesize carbohydrates from fatty acids Edit

The main fuel stored in the bodies of animals is fat. A young adult human's fat stores average between about 15– 20 kg, but varies greatly depending on age, gender, and individual disposition. [9] In contrast, the human body stores only about 400 g of glycogen, of which 300 g is locked inside the skeletal muscles and is unavailable to the body as a whole. The 100 g or so of glycogen stored in the liver is depleted within one day of starvation. [10] Thereafter the glucose that is released into the blood by the liver for general use by the body tissues, has to be synthesized from the glucogenic amino acids and a few other gluconeogenic substrates, which do not include fatty acids. [11]

Fatty acids are broken down to acetyl-CoA by means of beta oxidation inside the mitochondria, whereas fatty acids are synthesized from acetyl-CoA outside the mitochondrion, in the cytosol. The two pathways are distinct, not only in where they occur, but also in the reactions that occur, and the substrates that are used. The two pathways are mutually inhibitory, preventing the acetyl-CoA produced by beta-oxidation from entering the synthetic pathway via the acetyl-CoA carboxylase reaction. [11] It can also not be converted to pyruvate as the pyruvate decarboxylation reaction is irreversible. [10] Instead it condenses with oxaloacetate, to enter the citric acid cycle. During each turn of the cycle, two carbon atoms leave the cycle as CO2 in the decarboxylation reactions catalyzed by isocitrate dehydrogenase and alpha-ketoglutarate dehydrogenase. Thus each turn of the citric acid cycle oxidizes an acetyl-CoA unit while regenerating the oxaloacetate molecule with which the acetyl-CoA had originally combined to form citric acid. The decarboxylation reactions occur before malate is formed in the cycle. Malate is the only substance that can be removed from the mitochondrion to enter the gluconeogenic pathway to form glucose or glycogen in the liver or any other tissue. [11] There can therefore be no net conversion of fatty acids into glucose.

Only plants possess the enzymes to convert acetyl-CoA into oxaloacetate from which malate can be formed to ultimately be converted to glucose. [11]

Acetyl-CoA is formed into malonyl-CoA by acetyl-CoA carboxylase, at which point malonyl-CoA is destined to feed into the fatty acid synthesis pathway. Acetyl-CoA carboxylase is the point of regulation in saturated straight-chain fatty acid synthesis, and is subject to both phosphorylation and allosteric regulation. Regulation by phosphorylation occurs mostly in mammals, while allosteric regulation occurs in most organisms. Allosteric control occurs as feedback inhibition by palmitoyl-CoA and activation by citrate. When there are high levels of palmitoyl-CoA, the final product of saturated fatty acid synthesis, it allosterically inactivates acetyl-CoA carboxylase to prevent a build-up of fatty acids in cells. Citrate acts to activate acetyl-CoA carboxylase under high levels, because high levels indicate that there is enough acetyl-CoA to feed into the Krebs cycle and conserve energy. [12]

High plasma levels of insulin in the blood plasma (e.g. after meals) cause the dephosphorylation of acetyl-CoA carboxylase, thus promoting the formation of malonyl-CoA from acetyl-CoA, and consequently the conversion of carbohydrates into fatty acids, while epinephrine and glucagon (released into the blood during starvation and exercise) cause the phosphorylation of this enzyme, inhibiting lipogenesis in favor of fatty acid oxidation via beta-oxidation. [6] [8]

Unsaturated straight chain fatty acids Edit

Anaerobic desaturation Edit

Many bacteria use the anaerobic pathway for synthesizing unsaturated fatty acids. This pathway does not utilize oxygen and is dependent on enzymes to insert the double bond before elongation utilizing the normal fatty acid synthesis machinery. In Escherichia coli, this pathway is well understood.

  • FabA is a β-hydroxydecanoyl-ACP dehydrase – it is specific for the 10-carbon saturated fatty acid synthesis intermediate (β-hydroxydecanoyl-ACP).
  • FabA catalyzes the dehydration of β-hydroxydecanoyl-ACP, causing the release of water and insertion of the double bond between C7 and C8 counting from the methyl end. This creates the trans-2-decenoyl intermediate.
  • Either the trans-2-decenoyl intermediate can be shunted to the normal saturated fatty acid synthesis pathway by FabB, where the double bond will be hydrolyzed and the final product will be a saturated fatty acid, or FabA will catalyze the isomerization into the cis-3-decenoyl intermediate.
  • FabB is a β-ketoacyl-ACP synthase that elongates and channels intermediates into the mainstream fatty acid synthesis pathway. When FabB reacts with the cis-decenoyl intermediate, the final product after elongation will be an unsaturated fatty acid. [13]
  • The two main unsaturated fatty acids made are Palmitoleoyl-ACP (16:1ω7) and cis-vaccenoyl-ACP (18:1ω7). [14]

Most bacteria that undergo anaerobic desaturation contain homologues of FabA and FabB. [15] Clostridia are the main exception they have a novel enzyme, yet to be identified, that catalyzes the formation of the cis double bond. [14]

This pathway undergoes transcriptional regulation by FadR and FabR. FadR is the more extensively studied protein and has been attributed bifunctional characteristics. It acts as an activator of fabA and fabB transcription and as a repressor for the β-oxidation regulon. In contrast, FabR acts as a repressor for the transcription of fabA and fabB. [13]

Aerobic desaturation Edit

Aerobic desaturation is the most widespread pathway for the synthesis of unsaturated fatty acids. It is utilized in all eukaryotes and some prokaryotes. This pathway utilizes desaturases to synthesize unsaturated fatty acids from full-length saturated fatty acid substrates. [16] All desaturases require oxygen and ultimately consume NADH even though desaturation is an oxidative process. Desaturases are specific for the double bond they induce in the substrate. In Bacillus subtilis, the desaturase, Δ 5 -Des, is specific for inducing a cis-double bond at the Δ 5 position. [7] [16] Saccharomyces cerevisiae contains one desaturase, Ole1p, which induces the cis-double bond at Δ 9 . [7]

In mammals the aerobic desaturation is catalyzed by a complex of three membrane-bound enzymes (NADH-cytochrome b5 reductase, cytochrome b5, and a desaturase). These enzymes allow molecular oxygen, O2, to interact with the saturated fatty acyl-CoA chain, forming a double bond and two molecules of water, H2O. Two electrons come from NADH + H + and two from the single bond in the fatty acid chain. [6] These mammalian enzymes are, however, incapable of introducing double bonds at carbon atoms beyond C-9 in the fatty acid chain. [nb 1] .) Hence mammals cannot synthesize linoleate or linolenate (which have double bonds at the C-12 (= Δ 12 ), or the C-12 and C-15 (= Δ 12 and Δ 15 ) positions, respectively, as well as at the Δ 9 position), nor the polyunsaturated, 20-carbon arachidonic acid that is derived from linoleate. These are all termed essential fatty acids, meaning that they are required by the organism, but can only be supplied via the diet. (Arachidonic acid is the precursor the prostaglandins which fulfill a wide variety of functions as local hormones.) [6]

Odd-chain fatty acids Edit

Odd-chain fatty acids (OCFAs) are those fatty acids that contain an odd number of carbon atoms. The most common OCFAs are the saturated C15 and C17 derivatives, respectively pentadecanoic acid and heptadecanoic acid. [17] The synthesis of even-chained fatty acid synthesis is done by assembling acetyl-CoA precursors, however, propionyl-CoA instead of acetyl-CoA is used as the primer for the biosynthesis of long-chain fatty acids with an odd number of carbon atoms. [18]

Regulation In B. subtilis, this pathway is regulated by a two-component system: DesK and DesR. DesK is a membrane-associated kinase and DesR is a transcriptional regulator of the des gene. [7] [16] The regulation responds to temperature when there is a drop in temperature, this gene is upregulated. Unsaturated fatty acids increase the fluidity of the membrane and stabilize it under lower temperatures. DesK is the sensor protein that, when there is a decrease in temperature, will autophosphorylate. DesK-P will transfer its phosphoryl group to DesR. Two DesR-P proteins will dimerize and bind to the DNA promoters of the des gene and recruit RNA polymerase to begin transcription. [7] [16]

Pseudomonas aeruginosa

In general, both anaerobic and aerobic unsaturated fatty acid synthesis will not occur within the same system, however Pseudomonas aeruginosa and Vibrio ABE-1 are exceptions. [19] [20] [21] While P. aeruginosa undergoes primarily anaerobic desaturation, it also undergoes two aerobic pathways. One pathway utilizes a Δ 9 -desaturase (DesA) that catalyzes a double bond formation in membrane lipids. Another pathway uses two proteins, DesC and DesB, together to act as a Δ 9 -desaturase, which inserts a double bond into a saturated fatty acid-CoA molecule. This second pathway is regulated by repressor protein DesT. DesT is also a repressor of fabAB expression for anaerobic desaturation when in presence of exogenous unsaturated fatty acids. This functions to coordinate the expression of the two pathways within the organism. [20] [22]

Branched chain fatty acids are usually saturated and are found in two distinct families: the iso-series and anteiso-series. It has been found that Actinomycetales contain unique branch-chain fatty acid synthesis mechanisms, including that which forms tuberculosteric acid.


ATP Yield from Fatty Acid Oxidation

The amount of ATP obtained from fatty acid oxidation depends on the size of the fatty acid being oxidized. For our purposes here. we&rsquoll study palmitic acid, a saturated fatty acid with 16 carbon atoms, as a typical fatty acid in the human diet. Calculating its energy yield provides a model for determining the ATP yield of all other fatty acids.

The breakdown by an organism of 1 mol of palmitic acid requires 1 mol of ATP (for activation) and forms 8 mol of acetyl-CoA. Recall from Table 20.4.1 that each mole of acetyl-CoA metabolized by the citric acid cycle yields 10 mol of ATP. The complete degradation of 1 mol of palmitic acid requires the &beta-oxidation reactions to be repeated seven times. Thus, 7 mol of NADH and 7 mol of FADH2 are produced. Reoxidation of these compounds through respiration yields 2.5&ndash3 and 1.5&ndash2 mol of ATP, respectively. The energy calculations can be summarized as follows:

1 mol of ATP is split to AMP and 2Pi &minus2 ATP
8 mol of acetyl-CoA formed (8 × 12) 96 ATP
7 mol of FADH2 formed (7 × 2) 14 ATP
7 mol of NADH formed (7 × 3) 21 ATP
Total 129 ATP

The number of times &beta-oxidation is repeated for a fatty acid containing n carbon atoms is n/2 &ndash 1 because the final turn yields two acetyl-CoA molecules.

The combustion of 1 mol of palmitic acid releases a considerable amount of energy:

[C_<16>H_<32>O_2 + 23O_2 &rarr 16CO_2 + 16H_2O + 2,340 kcal]

The percentage of this energy that is conserved by the cell in the form of ATP is as follows:

The efficiency of fatty acid metabolism is comparable to that of carbohydrate metabolism, which we calculated to be 42%. For more information about the efficiency of fatty acid metabolism, see Section 20.5.

The oxidation of fatty acids produces large quantities of water. This water, which sustains migratory birds and animals (such as the camel) for long periods of time.


LIPIDS | Fatty Acids

Trivial Names and Systematic Nomenclature

Fatty acids are known by several names: volatile fatty acids from C 1 to C5, fatty acids from C6 to C24, long-chain fatty acids from C25 to C40, and very long chain fatty acids above C40. One can also find other names in the chemical nomenclature. The correct name for this group of carboxylic aliphatic acids (with or without any additional functions) is simply, fatty acids. It is possible to find trivial, Liege, and Geneva nomenclatures remaining in modern texts, despite the acceptance of the IUPAC nomenclature in 1957. Fatty acids are carboxylic aliphatic acids with the general formula, H(CH2)nCOOH. Aliphatic fatty acids are named by dropping the final ‘e’ from the parent alkane, and then adding the term ‘-oic acid’ to the root. The nomenclature for any additional functions on the principal chain follows the rules of the IUPAC convention of 1957. The first members in the homologous series are usually named using their trivial names ( Figure 1 ).

Figure 1 . Aliphatic saturated and monounsaturated fatty acids.

Fatty acids can have different chemical functional groups substituting for H- in the aliphatic principal chain, leading to different classes of fatty acids.


Biochemistry of Lipids, Lipoproteins and Membranes

Alfred H. Merrill Jr , Charles C. Sweeley , in New Comprehensive Biochemistry , 1996

3.1.1 Synthesis of the long-chain base backbone

The first step is the condensation of palmitoyl-CoA and L-serine, with loss of the carboxyl group of serine and production of 3-ketosphinganine ( Fig. 6 ). The reaction is catalyzed by the pyridoxal phosphate-dependent enzyme serine palmitoyltransferase and appears to be the rate-limiting step for sphingoid base biosynthesis. Serine palmitoyltransferase is highly selective for fatty acyl-CoA with 16 ± 1 carbon atoms, which accounts for the prevalence of long-chain bases of 18 carbon atoms (16 from palmitoyl-CoA and 2 from serine) in most sphingolipids [6] .

Robert Dickson and co-workers have isolated two genes (lcb1 and lcb2) that are required to overcome sphinganine auxotrophy in yeast with a defective serine palmitoyltransferase. Both genes are similar to δ-aminolevulinate synthase, which catalyzes the analogous condensation of glycine and succinyl-CoA. The reaction proceeds with overall retention of configuration of C2 of serine the likely mechanism is shown in Fig. 7 .

Fig. 7 . A probable reaction mechanism for serine palmitoyltransferase

(modified from [K. Krisnangkura, 1976].

As would be predicted from this mechanism, serine palmitoyltransferase undergoes time-dependent, irreversible (‘suicide’) inhibition by β-halo- l -alanines. Another inhibitor, l -cycloserine, has been shown to depress the level of central nervous system sphingolipids however, more potent and selective inhibitors have been recently isolated from microorganisms ( Table II ) ( Fig. 8 ). These include sphingofungins, lipoxamycins, and ISP-1 (also called myriocin). ISP-1 is a powerful immunosuppressive agent, being one to two orders of magnitude more potent than cyclosporin A in inhibition of proliferation in the mouse allogenic mixed lymphocyte reaction, and generation of allo-reactive cytotoxic T lymphocytes [20] . ISP-1 resembles a transition state intermediate of serine palmitoyltransferase (cf. Figs. 7 and 8 ), and inhibits this enzyme with Ki < 1 nM.

Fig. 8 . Inhibitors of sphingolipid biosynthesis that have been isolated from microorganisms. For references for these compounds, see Table II .

Sphingoid base synthesis is regulated by the availability of the precursors of this pathway [6] furthermore, addition of lipoproteins or free sphingoid bases to cells in culture reduces de novo sphingolipid biosynthesis, perhaps by transcriptional down-regulation of serine palmitoyltransferase [G. van Echten, 1990].

The next step of sphingoid base synthesis is the reduction of 3-keto-sphinganine ( Fig. 6 ) by the transfer of the α-hydrogen of NADPH to C3 of the long-chain base. This reaction is rapid because the 3-keto intermediate is not seen in cells or in vitro assays if NADPH is available.


Oxidation of Fatty Acids

Fatty acids stored in the form of TAG in the adipose tissue serve as the main energy reserve of the human body. They are released from these stores into the blood to be carried to the tissues for oxidation. Oxidation of fatty acids in tissues generates a tremendous amount of energy. This energy can be used in various needed within the cells.

Mobilization of fatty acids

The stored fatty acids must be first mobilized to be used for obtaining energy. The mobilization of fatty acids is done by specific lipases. These are the enzymes that cleave fatty acids from glycerol and release them into the blood. Important lipases are

  • Adipose triacylglycerol lipase (ATGL), acts on triacylglycerol.
  • Hormone-sensitive lipase (HSL), acts on diacylglycerol.
  • Monoacylglycerol lipase (MAGL), acts on monoacylglycerol.

Once the fatty acids have been released, they cross the plasma membrane to enter the blood where they bind the albumin protein present in plasma. Albumin carries these fatty acids to various tissues where they can be oxidized to generate energy. Two types of cells cannot use fatty acids as an energy source

  • Red blood cells, because they lack mitochondria and cannot oxidize fatty acids.
  • Brain cells, because fatty acids cannot cross the blood-brain barrier.

Glycerol is left behind in the adipose tissue after the mobilization of fatty acids. Adipose tissues cannot metabolize glycerol any further because they don’t have the glycerol kinase enzyme. The glycerol molecule is also released into the blood to be carried to the liver where it can be used for making new triglycerides or can be consumed in the glycolytic pathway.

Beta-Oxidation

Fatty acids release energy when they are broken down in the presence of oxygen. Oxidation of fatty acids takes place within the mitochondria. There are different types of fatty acid oxidation, but the most common is beta-oxidation.

In beta-oxidation of fatty acids, two carbon atoms are released from fatty acids in one chain of reactions. It involves the following steps

Transport of Fatty acids into Mitochondria

Beta-oxidation takes place inside the mitochondria of cells. Small and medium-chain fatty acids can easily cross the mitochondrial membrane however, long-chain fatty acids cannot cross it. They need special carriers to be transported into the mitochondria. This is done by carnitine and the process is called carnitine shuttle.

Once inside the mitochondria, the fatty acids are activated by enzymes of the matrix into their CoA derivatives. These CoA derivatives then undergo beta-oxidation.

Reactions of Oxidation

Each fatty acyl CoA undergoes following four steps of beta-oxidation

  1. Fatty acyl CoA dehydrogenase introduces a double bond between 2 nd and 3 rd carbon from the CoA end of fatty acyl CoA and converts it into trans-2-Enoyl CoA. One FAD is reduced to FADH2 in this process.
  2. Hydroxylation of the enoyl group is carried out by 2,3-Enoyl CoA hydrolase enzyme forming a 3-hydroxyacyl CoA.
  3. The enzyme 3-hydroxyacyl CoA dehydrogenase oxidizes the above product resulting in the formation of 3-ketoacyl CoA. One NAD + molecule is reduced to NADH2 in this reaction.
  4. 3-Ketoacyl CoA thiolase removes the acetyl CoA from the terminal end leaving behind the fatty acyl CoA that is two carbon deficient from the original molecule.

The acetyl CoA released in the fourth step undergoes further oxidation in the citric acid cycle.

The above four steps are repeated until the acetyl CoA is left behind.

The net product of one cycle of beta-oxidation is one NADH2, one FADH2, and one acetyl CoA.

With this, we conclude our discussion on fatty acids and triglycerides.


Fats in the Diet

Fats are necessary to maintain the body. Dietary fats exist mainly in the form of triglycerides. Fats are classified as high-energy food. They provide about 9 kilocalorie (kcal) of energy per gram consumed. In contrast, both carbohydrates and proteins provide about 4 kcal of energy per gram consumed. As a result, fat is the most efficient way to store energy. If one consumes more calories than he or she needs, some of the excess calorie source is converted to fat.

It is generally recommended that not more than 30 percent of one's dietary calories should derive from fat (and of this 30 percent, 10 percent should be monounsaturated, and 10 percent, polyunsaturated). It appears that consumption of greater amounts of saturated fats (compared to amounts of monounsaturated and polyunsaturated fats) is related to higher levels of cholesterol in the blood and a greater risk of heart disease. In the United States the average diet is about 34 percent fat and (13 percent saturated fat).


Abstract

Lipid metabolism, in particular the synthesis of fatty acids (FAs), is an essential cellular process that converts nutrients into metabolic intermediates for membrane biosynthesis, energy storage and the generation of signalling molecules. This Review explores how different aspects of FA synthesis promote tumorigenesis and tumour progression. FA synthesis has received substantial attention as a potential target for cancer therapy, but strategies to target this process have not yet translated into clinical practice. Furthermore, efforts to target this pathway must consider the influence of the tumour microenvironment.


Fatty acids

Figure 2.190 - Saturated fatty acid (stearic acid) and unsaturated fatty acid (oleic acid)

The most ubiquitous lipids in cells are the fatty acids. Found in fats, glycerophospholipids, sphingolipids and serving as as membrane anchors for proteins and other biomolecules, fatty acids are important for energy storage, membrane structure, and as precursors of most classes of lipids. Fatty acids, as can be seen from Figure 2.190 are characterized by a polar head group and a long hydrocarbon tail. Fatty acids with hydrocarbon tails that lack any double bonds are described as saturated, while those with one or more double bonds in their tails are known as unsaturated fatty acids. The effect of double bonds on the fatty acid tail is to introduce a kink, or bend, in the tail, as shown for oleic acid.

Figure 2.191 - Arachidonic acid - A polyunsaturated fatty acid Wikipedia

Stearic acid, a saturated fatty acid, by contrast has a straight hydrocarbon tail. Figures 2.190-2.194 show the most common saturated and unsaturated fatty acids. Fatty acids with unsaturated tails have a lower melting temperature than those with saturated tails of the same length. Shorter tails also decrease melting temperature. These properties carry over to the fats/oils containing them.

Figure 2.192 - Saturated fatty acids. Number of carbons in right column Wikipedia

Fatty acids with more than one double bond are called polyunsaturated. Plants are excellent sources of unsaturated and polyunsaturated fatty acids. The position of the double bond(s) in fatty acids has important considerations both for their synthesis and for their actions in the body. Biochemically, the double bonds found in fatty acids are predominantly in the cis configuration. So-called trans fats arise as a chemical by-product of partial hydrogenation of vegetable oil.

Figure 2.193 - Unsaturated fatty acids. Right column Indicates number of carbons and double bonds Wikipedia

In humans, consumption of trans fat raises low density lipoprotein (LDL) levels and lowers high density lipoprotein (HDL) levels. Each is thought to contribute to the risk of developing coronary artery disease. The most Figure 2.194 - Fatty acid models. Carboxyl end labeled in red Wikipedia common fatty acids in our body include palmitate, stearate, oleate, linolenate, linoleate, and arachidonate. Two notable shorter fatty acids are nonanoic (9 carbons) and decanoic acid (10 carbons), both of which appear to have anti-seizure effects. Decanoic acid directly inhibits excitatory neurotransmission in the brain and may contribute to the anticonvulsant effect of the ketogenic diet.

Figure 2.194 - Fatty acid models. Carboxyl end labeled in red Wikipedia

Figure 2.195 shows two different systems for locating double bonds in a fatty acid. The &omega system counts carbons starting with the methyl end (shown in red) while the &Delta system counts from the carboxyl end (shown in blue). For example, an &omega-3 (omega 3) fatty acid would have a double bond at the third carbon from the methyl end. In the &Delta system, a fatty acid that has a cis double bond at carbon 6, counting from the carboxyl end, would be written as cis-&Delta6.

Figure 2.195 - &Delta and &omega numbering systems for fatty acids Image by Pehr Jacobson

Fatty acids are described as essential fatty acids if they must be in the diet (can&rsquot be synthesized by the organism) and nonessential fatty acids if the organism can synthesize them. Humans and other animals lack the desaturase enzymes necessary to make double bonds at positions greater than &Delta-9, so fatty acids with double bonds beyond this position must be obtained in the diet. Linoleic acid and linolenic acid, both fall in this category. Related unsaturated fatty acids can be made from these fatty acids, so the presence of linoleic and linolenic acids in the diet eliminates the need to have all unsaturated fatty acids in the diet. Both linoleic and linolenic acid contain 18 carbons, but linoleic acid is an &omega-6 fatty acid, whereas linolenic acid is an &omega-3 fatty acid. Notably, &omega-6 fatty acids tend to be proinflammatory, whereas &omega-3 fatty acids are lesser so.

Figure 2.196 - Structure of a fat/oil

Fats and oils are the primary energy storage forms of animals and are also known as triacylglycerols and triglycerides, since they consist of a glycerol molecule linked via ester bonds to three fatty acids (Figure 2.196). Fats and oils have the same basic structure. We give the name fat to those compounds that are solid at room temperature and the name oil to those that are liquid at room temperature. Note that biological oils are not the same as petroleum oils.

Increasing the number of unsaturated fatty acids (and the amount of unsaturation in a given fatty acid) in a fat decreases the melting temperature of it. Organisms like fish, which live in cool environments, have fats with more unsaturation and this is why fish oil contains polyunsaturated fatty acids.

Figure 2.197 - Lipase action on a fat Image by Aleia Kim

Fats are stored in the body in specialized cells known as adipocytes. Enzymes known as lipases release fatty acids from fats by hydrolysis reactions (Figure 2.197). Triacylglycercol lipase (pancreatic - Figure 2.198) is able to cleave the first two fatty acids from the fat. A second enzyme, monoacylglycerol lipase, cleaves the last fatty acid. Fats can be synthesized by replacing the phosphate on phosphatidic acid with a fatty acid.

Figure 2.198 - Pancreatic lipase

Glycerophospholipids

Glycerophospholipids (phosphoglycerides) are important components of the lipid bilayer of cellular membranes. Phosphoglycerides are structurally related to fats, as both are derived from phosphatidic acid (Figure 2.199). Phosphatidic acid is a simple glycerophospholipid that is usually converted into phosphatidyl compounds. These are made by esterifying various groups, such as ethanolamine, serine, choline, inositol, and others (Figure 2.200) to the phosphate of phosphatidic acid. All of these compounds form lipid bilayers in aqueous solution , due to their amphiphilic nature.

Figure 2.199 - Structure of phosphatidic acid. R1 and R2 are alkyl groups of fatty acids.

Phosphatidylethanolamines

Since all glycerolipids can have a variety of fatty acids at positions 1 and 2 on the glycerol, they all are families of compounds. The phosphatidylethanolamines are found in all living cells and are one of the most common phosphatides, making up about 25% of them. They are common constituents of brain tissue and in the spinal cord, making up as much as 45% of the total phospholipids. Phosphatidylethanolamines are asymmetrically distributed across membranes, being preferentially located on the inner leaflet (closest to the cytoplasm) of the plasma membrane. Metabolically, phosphatidylethanloamines are precursors of phosphatidylcholines. Phosphatidylserines Phosphatidylserines are another group of phosphatidyl compounds that are preferentially distributed across the lipid bilayer of the plasma membrane. Like the phosphatidylethanolamines, phosphatidylserines are preferentially located on the inner leaflet of the plasma membrane. When apoptosis (cell suicide) occurs, the preferential distribution is lost and the phosphatidylserines appear on the outer leaflet where they serve as a signal to macrophages to bind and destroy the cell.

Figure 2.200 - Four common components of phosphatides Wikipedia

Phosphatidylcholines

Phosphatidylcholines (Figure 2.201) are another group of important membrane components. They tend to be found more commonly on the outer leaflet of the plasma membrane. Nutritionally, the compounds are readily obtained from eggs and soybeans. Phosphatidylcholines are moved across membranes by Phosphatidylcholine transfer protein (PCTP). This protein, which is sensitive to the levels of phosphatidylcholines, acts to stimulate the activity of a thioesterase (breaks thioester bonds, such as acyl-CoAs) and activates PAX3 transcription factors.

Figure 2.201 - Phosphatidylcholine

Cardiolipins

Cardiolipins are an unusual set of glycerophospholipids in containing two diacylglycerol backbones joined in the middle by a diphosphoglycerol (Figure 2.202). It is an important membrane lipid, constituting about 20% of the inner mitochondrial membrane and is found in organisms from bacteria to humans. In both plants and animals, it is found almost totally in the inner mitochondrial membrane.

Figure 2.202 - Cardiolipin

The molecules appear to be required for both Complex IV and Complex III of the electron transport chain to maintain its structure. The ATP synthase enzyme (Complex V) of the oxidative phosphorylation system also binds four molecules of cardiolipin. It has been proposed that cardiolipin functions as a proton trap in the process of proton pumping by Complex IV.

Figure 2.203 - Cardiolipin oxidation and the release of cytochrome C in apoptosis

Cardiolipin also plays a role in apoptosis. As shown in Figure 2.203, oxidation of cardiolipin by a cardiolipin-specific oxygenase causes cardiolipin to move from the inner mitochondrial membrane to the outer one, helping to form a permeable pore and facilitating the transport of cytochrome c out of the intermembrane space and into the cytoplasm - a step in the process of apoptosis.

Figure 2.204 - Diacylglycerol

Diacylglycerol

Diacylglycerol (also called diglyceride and DAG - Figure 2.204) is an important intermediate in metabolic pathways. It is produced, for example, in the first step of the hydrolysis of fat and is also produced when membrane lipids, such as PIP2 (phosphatidylinositol-4,5-bisphosphate) are hydrolyzed by phospholipase C in a signaling cascade.

DAG is itself a signaling compound, binding to protein kinase C to activate it to phosphorylate substrates. Synthesis of DAG begins with glycerol-3-phosphate, which gains two fatty acids from two acyl-CoAs to form phosphatidic acid. Dephosphorylation of phosphatidic acid produces DAG. DAG can also be rephosphorylated by DAG kinase to re-make phosphatidic acid or another fatty acid can be added to make fat.

Figure 2.205 - Inositol

Though technically not a lipid itself, inositol is found in many lipids. Inositol is a derivative of cyclohexane containing six hydroxyl groups - one on each carbon (Figure 2.205. It has nine different stereoisomers of which one, cis-1,2,3,5-trans-4,6- cyclohexanehexol (called myo-inositol) is the most common. It has a sweet taste (half that of sucrose).

Figure 2.206 - Phytic acid

Numerous phosphorylated forms of the compound exist, from a single phosphate to six (one on each carbon). Phytic acid, for example, in plants, has six phosphates (Figure 2.206) that it uses to store phosphate. Inositol is produced from glucose and was once considered vitamin B8, but is made by the body in adequate amounts, so it is not now considered a vitamin. Phosphorylated forms of inositol are found in phosphoinositides, such as PIP2 and PIP3, both of which are important in signaling processes. Some of these include insulin signaling, fat catabolism, calcium regulation, and assembly of the cytoskeleton.

Phosphoinositides

Figure 2.207 - Structure of PIP2

Compounds based on phosphatidylinositol (PI) are often called phosphoinositides. These compounds have important roles in signaling and membrane trafficking. Hydroxyls on carbons 3,4, and 5 of the inositol ring are targets for phosphorylation by a variety of kinases. Seven different combinations are used. Steric hindrance inhibits phosphorylation of carbons 2 or 6. Naming of these phosphorylated compounds follows generally as PI(#P)P, PI(#P, #P)P, or PI(#P, #P, #P)P where #P refers to the number of the carbon where a phosphate is located. For example, PI(3)P refers to a phosphatidyl compound with a phosphate added to carbons 3 of the inositol ring, whereas PI(3,4,5)P is a phosphatidyl compound with a phosphate added to carbons 3,4,and 5.

Phosphatidylinositol-4,5- bisphosphate

Figure 2.208 - Phosphatidylinositol-4- phosphate

Phosphatidylinositol-4,5-bisphosphate (PIP2 - Figure 2.207) is a phospholipid of plasma membranes that functions in the phospholipase C signaling cascade. In this signaling pathway, hydrolysis catalyzed by phospholipase C releases inositol-1,4,5- trisphosphate (IP3) and diacylglycerol. Synthesis of PIP2 begins with phosphatidylinositol, which is phosphorylated at position 4 followed by phosphorylation at position 5 by specific kinases.

PIP2 can be phosphorylated to form the signaling molecule known as phosphatidylinositol (3,4,5)-trisphosphate (PIP3). Along with PIP3, PIP2 serves as a docking phospholipid for the recruitment of proteins that play roles in signaling cascades. Binding of PIP2 is also required by inwardly directed potassium channels.

Phosphatidylinositol (3,4,5)- trisphosphate

Phosphatidylinositol (3,4,5)-trisphosphate (PIP3) is an important molecule for the activation of signaling proteins, such as AKT, which activates anabolic signaling pathways related to growth and survival. PIP3 can be dephosphorylated by phosphatase PTEN to yield PIP2 and can be synthesized from PIP2 by kinase action of Class I PI 3- kinases. Kinase activity to synthesize PIP3 results in movement of PIP3-binding proteins to the plasma membrane. They include Akt/ PKB, PDK1, Btk1, and ARNO and each is activated by binding to PIP3.

Plasmalogens

Figure 2.209 - Plasmalogen - A vinyl ether lipid Wikipedia

A special class of the glycerophospholipids are the plasmalogens (Figure 2.209). They differ in containing a vinyl ether linkage at position 1 of glycerol, in contrast to other glycerophopsholipids, which have an ester linkage at this position. Position 2 of each is an ester. The precursor for the ether linkage is typically a 16 or 18 carbon saturated alcohol or an 18 carbon unsaturated alcohol.

At the phosphate tail, the most commonly attached groups are ethanolamine or choline. Plasmalogens are found abundantly in humans in heart (30-40% of choline phospholipids). 30% of the glycerophospholipids in brain are plasmalogens and 70% of the ethanolamine lipids of the myelin sheath of nerve cells are plasmalogens.

Though their function is not understood, it is believed that plasmalogens may provide some protection against reactive oxygen species and have roles in signaling.

Lecithin is a generic term for a combination of lipid substances that includes phosphoric acid, glycerol, glycolipids, triglycerides, and phospholipids. Lecithin is a wetting agent helpful with emulsification and encapsulation and is even used as an anti-sludge additive in motor lubricants. Lecithin is used in candy bars to keep cocoa and cocoa butter from separating. Though considered safe as a food ingredient, lecithin can be converted by gut bacteria to trimethylamine-N-oxide which may contribute to cholesterol deposition and atherosclerosis.

Sphingolipids

Figure 2.210 - Sphingosine and a ceramide made from it Wikipedia

Fatty acids are also components of a broad class of molecules called sphingolipids. Sphingolipids are structurally similar to glycerophospholipids, though they are synthesized completely independently of them starting with palmitic acid and the amino acid serine. Sphingolipids are named for the amino alcohol known as sphingosine (Figure 2.210), though they are not directly synthesized from it. Figure 2.211 shows the generalized structure of sphingolipids.

Figure 2.211 Schematic structure of a sphingolipid

If the R-group is a hydrogen, the molecule is called a ceramide. When the R-group is phosphoethanolamine the resulting molecule is sphingomyelin, an important component of the myelin sheath and lipid membranes. If a single, simple sugar is instead added, a cerebroside is created (Figure 2.212). Addition of a complex oligosaccharide creates a ganglioside.

Complex sphingolipids may play roles in cellular recognition and signaling. Sphingolipids are found most abundantly in plasma membrane and are almost completely absent from mitochondrial and endoplasmic reticulum membranes. In animals, dietary sphingolipids have been linked to reduced colon cancer, reductions in LDLs, and increases in HDLs. Like the glycerophospholipids, sphingolipids are amphiphilic. Most sphingolipids except sphingomyelin do not contain phosphate.

Figure 2.212 - Categories of Sphingolipid Wikipedia

Eicosanoids

Figure 2.213 - Arachidonic acid drawn as straight (top) and bent (bottom)

Fatty acids made from omega-6 and omega-3 fatty acids include three important fatty acids containing 20 carbons. They include arachidonic acid (an &omega-6 fatty acid with four double bonds (&Delta-5,8,11,14) - Figure 2.213), eicosapentaenoic acid (an &omega-3 fatty acid with five double bonds, and dihomo-&gamma-linolenic acid (an &omega-6 fatty acid with three double bonds). The class of compounds known as eicosanoids is made by oxidation of these compounds. Subclasses include include prostaglandins, prostacyclins, thromboxanes, lipoxins, leukotrienes, and endocannabinoids (Figures 2.214-2.219). Eicosanoids play important roles affecting inflammation, immunity, mood, and behavior.

Prostaglandins

Figure 2.214 - Prostaglandin PGH2

A collection of molecules acting like hormones, prostaglandins are derived from arachidonic acid and have many differing (even conflicting) physiological effects. These include constriction or dilation of vascular smooth muscle cells, induction of labor, regulation of inflammation, and action on the thermoregulatory center of the hypothalamus to induce fever, among others.

Prostaglandins are grouped with the thromboxanes (below) and prostacyclins (below), as prostanoids. The prostanoids, which all contain 20 carbons are a subclass of the eicosanoids. Prostaglandins are found in most tissues of higher organisms. They are autocrine or paracrine compounds produced from essential fatty acids. The primary precursor of the prostaglandins is the fatty acid known as arachidonic acid and the prostaglandin made from it is known as PGH2 (Figure 2.214), which, in turn is a precursor of other prostaglandins, as well as the prostacyclins and thromboxanes.

Interesting prostaglandins

PGD2 - inhibits hair follicle growth, vasodilator, causes bronchial constriction, higher in lungs of asthmatics than others.

Figure 2.215 Prostaglandin E

PGE2 (Figure 2.215) - exerts effects in labor (soften cervix, uterine contraction), stimulates bone resorption by osteoclasts, induces fever, suppresses T-cell receptor signaling, vasodilator, inhibits release of noradrenalin from sympathetic nerve terminals. It is a potent activator of the Wnt signaling pathway.

A prostaglandin can have opposite effects, depending on which receptor it binds to. Binding of PGE2 to the EP1 receptor causes bronchoconstriction and smooth muscle contraction, whereas binding of the same molecule to the EP2 receptor causes bronchodilation and smooth muscle relaxation.

Figure 2.216 - Prostaglandin F2&alpha

PGF2&alpha (Figure 2.216)- uterine contractions, induces labor, bronchoconstriction.

PGI2 - vasodilation, bronchodilation, inhibition of platelet aggregation.

Thromboxanes

Figure 2.217 Thromboxane A2 2

Thromboxanes play roles in clot formation and named for their role in thrombosis. They are potent vasoconstrictors and facilitate platelet aggregation. They are synthesized in platelets, as well. The anti-clotting effects of aspirin have their roots in the inhibition of synthesis of PGH2, which is the precursor of the thromboxanes. The most common thromboxanes are A2 (Figure 2.217) and B2.

Prostacyclin

Figure 2.218 - Prostacyclin

Prostacyclin (also known as prostaglandin I2 or PGI2 - Figure 2.218) counters the effects of thromboxanes, inhibiting platelet activation and acting as vasodilators. It is produced from PGH2 by action of the enzyme prostacyclin synthase.

Leukotrienes

Figure 2.219 - Leukotriene A4 (LTA4)


Another group of eicosanoid compounds are the leukotrienes (Figure 2.219). Like prostaglandins, leukotrienes are made from arachidonic acid. The enzyme catalyzing their formation is a dioxygenase known as arachidonate 5-lipoxygenase. Leukotrienes are involved in regulating immune responses. They are found in leukocytes and other immunocompetent cells, such as neutrophils, monocytes, mast cells, eosinophils, and basophils. Leukotrienes are associated with production of histamines and prostaglandins, which act as mediators of inflammation. Leukotrienes also trigger contractions in the smooth muscles of the bronchioles. When overproduced, they may pay a role in asthma and allergic reactions. Some treatments for asthma aim at inhibiting production or action of leukotrienes.

Cholesterol

Figure 2.220 - Structure of cholesterol

Arguably, no single biomolecule has generated as much discussion and interest as has cholesterol (Figure 2.220). Certainly, from the perspective of the Nobel Prize committee, no small molecule even comes close, with 13 people having been awarded prizes for work on it. Evidence for cholesterol&rsquos importance comes from the study of brain tissue where it comprises 10-15% of the dry mass.

Membrane flexibility

Figure 2.221 - Sitosterol - A phytosterol

In animal cells, cholesterol provides for membrane flexibility that allows for cellular movement that is in contrast to plant and bacterial cells with fixed structures. Cholesterol is made in many cells of the body, with the liver making the greatest amount. The anabolic pathway leading to synthesis of cholesterol is known as the isoprenoid pathway and branches of it lead to other molecules including other fat-soluble vitamins.

Figure 2.222 - Margarine - A common source of trans fat Wikipedia

Cholesterol is only rarely found in prokaryotes (Mycoplasma, which requires it for growth, is an exception) and is found in only trace amounts in plants. Instead, plants synthesize similar compounds called phytosterols (Figure 2.221). On average, the body of a 150 pound adult male makes about 1 gram of cholesterol per day, with a total content of about 35 grams.

Figure 2.223 - Cholesterol - Ball and stick model

Cholesterol&rsquos (and other lipids&rsquo) hydrophobicity requires special packaging into lipoprotein complexes (called chylomicrons, VLDLs, IDLs, LDLs, and HDLs) for movement in the lymph system and bloodstream. Though cholesterol can be made by cells, they also take it up from the blood supply by absorbing cholesterol-containing LDLs directly in a process called receptor-mediated endocytosis.

Oxidative damage to LDLs can lead to formation of atherosclerotic plaques and this is why cholesterol has gotten such a negative image in the public eye. The liver excretes cholesterol through the bile for elimination into the digestive system, but the compound is recycled there.

Reducing cholesterol levels

Figure 2.224 - Ezetimibe - An inhibitor of cholesterol absorption

Strategies for reducing cholesterol in the body focus primarily on three areas - reducing consumption, reducing endogenous synthesis, and reducing the recycling. Dietary considerations, such as saturated fat versus unsaturated fat consumption are currently debated. Dietary trans fats, though, correlate with incidence of coronary heart disease. Consumption of vegetables may provide some assistance with reducing levels of cholesterol recycled in the digestive system, because plant phytosterols compete with cholesterol for reabsorption and when this happens, a greater percentage of cholesterol exits the body in the feces. Drugs related to penicillin are also used to inhibit cholesterol recycling. One of these is ezetimibe, shown in Figure 2.224.

Figure 2.225 - All-trans retinol

Genetic defects in the cholesterol movement system are a cause of the rare disease known as familial hypercholesterolemia in which the blood of afflicted individuals contains dangerously high levels of LDLs. Left untreated, the disease is often fatal in the first 10-20 years of life. While LDLs have received (and deserve) a bad rap, another group of lipoprotein complexes known as the HDLs (high density lipoprotein complexes) are known as &ldquogood cholesterol&rdquo because their levels correlate with removal of debris (including cholesterol) from arteries and reduce inflammation.

Membrane function

In membranes, cholesterol is important as an insulator for the transmission of signals in nerve tissue and it helps to manage fluidity of membranes over a wide range of temperatures. Stacked in the lipid bilayer, cholesterol decreases a membrane&rsquos fluidity and its permeability to neutral compounds, as well as protons and sodium ions. Cholesterol may play a role in signaling by helping with construction of lipid rafts within the cell membrane.

Figure 2.226 - 11-cis retinal

Vitamin A comes in three primary chemical forms, retinol (storage in liver - Figure 2.225), retinal (role in vision - Figure 2.226), and retinoic acid (roles in growth and development). All vitamin A forms are diterpenoids and differ only in the chemical form of the terminal group. Retinol is mostly used as the storage form of the vitamin.

Retinol is commonly esterified to a fatty acid and kept in the liver. In high levels, the compound is toxic. Retinol is obtained in the body by hydrolysis of the ester or by reduction of retinal. Importantly, neither retinal nor retinol can be made from retinoic acid. Retinoic acid is important for healthy skin and teeth, as well as bone growth. It acts in differentiation of stem cells through a specific cellular retinoic acid receptor.

Figure 2.227 - &beta-Carotene

Good sources of vitamin A are liver and eggs, as well as many plants, including carrots, which have a precursor, &beta-carotene (Figure 2.227) from which retinol may be made by action of a dioxygenase.

Light sensitivity The conjugated double bond system in the side chain of vitamin A is sensitive to light and can flip between cis and trans forms on exposure to it. It is this response to light that makes it possible for retinal to have a role in vision in the rods and cones of the eyes. Here, the aldehyde form (retinal) is bound to the protein rhodopsin in the membranes of rod and cone cells.

Figure 2.228 - Color sensitivity for cones and rods Image by Aleia Kim

When exposed to light of a particular wavelength, the &ldquotail&rdquo of the retinal molecule will flip back and forth from cis to trans at the double bond at position 11 of the molecule. When this happens, a nerve signal is generated that signals the brain of exposure to light. Slightly different forms of rhodopsin have different maximum absorption maxima allowing the brain to perceive red, green and blue specifically and to assemble those into the images we see (Figure 2.228). Cones are the cells responsible for color vision, whereas rods are mostly involved in light detection in low light circumstances.

Deficiency and surplus

Deficiency of vitamin A is common in developing countries and was inspiration for the design and synthesis of the geneticallymodified golden rice, which is used as a source of vitamin A to help prevent blindness in children. Overdose of vitamin A, called hypervitaminosis A is dangerous and can be fatal. Excess vitamin A is also suspected to be linked to osteoporosis. In smokers, excess vitamin A is linked to an increased rate of lung cancer, but non-smokers have a reduced rate.

Vitamin D

Figure 2.229 - Cholecalciferol - Vitamin D3

The active form of vitamin D plays important roles in the intestinal absorption of calcium and phosphate and thus in healthy bones. Technically, vitamin D isn&rsquot even a vitamin, as it is a compound made by the body. Rather, it behaves more like a hormone.

Derived from ultimately from cholesterol, vitamin D can be made in a reaction catalyzed by ultraviolet light. In the reaction, the intermediate 7-dehydrocholesterol is converted to cholecalciferol (vitamin D3) by the uv light (Figure 2.229). The reaction occurs most readily in the bottom two layers of the skin shown in Figure 2.230.

Figure 2.230 - Layers of the skin. Outside is at top.

Forms of vitamin D

Five different compounds are referred to as vitamin D. They are

Vitamin D1 - A mixture of ergocalciferol and lumisterol

Vitamin D2 - Ergocalciferol

Vitamin D3 - Cholecalciferol Vitamin

D4 - 22-Dihydroergocalciferol Vitamin

Vitamin D3 is the most common form used in vitamin supplements and it and vitamin D2 are commonly obtained in the diet, as well. The active form of vitamin D, calcitriol (Figure 2.231), is made in the body in controlled amounts. This proceeds through two steps from cholecalciferol. First, a hydroxylation in the liver produces calcidiol and a second hydroxylation in the kidney produces calcitriol. Monocyte macrophages can also synthesize vitamin D and they use is as a cytokine to stimulate the innate immune system.

Mechanism of action

Calcitriol moves in the body bound to a vitamin D binding protein, which delivers it to target organs. Calcitriol inside of cells acts by binding a vitamin D receptor (VDR), which results in most of the vitamin&rsquos physiological effects. After binding calcitriol, the VDR migrates to the nucleus where it acts as a transcription factor to control levels of expression of calcium transport proteins (for example) in the intestine. Most tissues respond to VDR bound to calcitriol and the result is moderation of calcium and phosphate levels in cells.

Deficiency/excess

Figure 2.231 - Calcitriol - Active form of vitamin D

Deficiency of vitamin D is a cause of the disease known as rickets, which is characterized by soft, weak bones and most commonly is found in children. It is not common in the developed world, but elsewhere is of increasing concern.

Excess of vitamin D is rare, but has toxic effects, including hypercalcemia, which results in painful calcium deposits in major organs. Indications of vitamin D toxicity are increased urination and thirst. Vitamin D toxicity can lead to mental retardation and many other serious health problems.

Figure 2.232 &alpha-tocopherol - The most biologically active form of vitamin E Figure

Vitamin E comprises a group of two compounds (tocopherols and tocotrienols - Figure 2.232) and stereoisomers of each. It is commonly found in plant oils. The compounds act in cells as fat-soluble antioxidants. &alpha-tocopherol (Figure 2.233), the most active form of the vitamin, works 1) through the glutathione peroxidase protective system and 2) in membranes to interrupt lipid peroxidation chain reactions. In both actions, vitamin E reduces levels of reactive oxygen species in cells.

Figure 2.233 - Structure of tocotrienols

Figure 2.234 - Lipid peroxidation reactions

Vitamin E scavenges oxygen radicals (possessing unpaired electrons) by reacting with them to produce a tocopheryl radical. This vitamin E radical can be converted back to its original form by a hydrogen donor. Vitamin C is one such donor. Acting in this way, Vitamin E helps reduce oxidation of easily oxidized compounds in the lipid peroxidation reactions (Figure 2.234).

Vitamin E also can affect enzyme activity. The compound can inhibit action of protein kinase C in smooth muscle and simultaneously activate catalysis of protein phosphatase 2A to remove phosphates, stopping smooth muscle growth.

Deficiency/excess

Deficiency of vitamin E can lead to poor conduction of nerve signals and other issues arising from nerve problems. Low levels of the vitamin may be a factor in low birth weights and premature deliveries. Deficiency, however, is rare, and not usually associated with diet.

Excess Vitamin E reduces vitamin K levels, thus reducing the ability to clot blood. Hypervitaminosis of vitamin E in conjunction with aspirin can be life threatening. At lower levels, vitamin E may serve a preventative role with respect to atherosclerosis by reducing oxidation of LDLs, a step in plaque formation.

Figure 2.235 - Forms of vitamin K Image by Pehr Jacobson

Like the other fat-soluble vitamins, Vitamin K comes in multiple forms (Figure 2.235) and is stored in fat tissue in the body. There are two primary forms of the vitamin - K1 and K2 and the latter has multiple sub-forms . Vitamins K3, K4, and K5 are made synthetically, not biologically.

Figure 2.236 - Recycling of vitamin K Wikipedia

Vitamin K is used as a co-factor for enzymes that add carboxyl groups to glutamate side chains of proteins to increase their affinity for calcium. Sixteen such proteins are known in humans. They include proteins involved in blood clotting (prothrombin (called Factor II), Factors VII, IX, and X), bone metabolism (osteocalcin, also called bone Gla protein (BGP), matrix Gla protein (MGP), and periostin) and others.

Modification of prothrombin is an important step in the process of blood clotting (see HERE). Reduced levels of vitamin K result in less blood clotting, a phenomenon sometimes referred to as blood thinning. Drugs that block recycling of vitamin K (Figure 2.236) by inhibiting the vitamin K epoxide reductase, produce lower levels of the vitamin and are employed in treatments for people prone to excessive clotting. Warfarin (coumadin) is one such compound that acts in this way and is used therapeutically. Individuals respond to the drug differentially, requiring them to periodically be tested for levels of clotting they possess, lest too much or too little occur.

Figure 2.237 - Steroid numbering scheme Image by Pehr Jacobson

Vitamin K1 is a stereoisomer of the plant photosystem I electron receptor known as phylloquinone and is found abundantly in green, leafy vegetables. Phylloquinone is one source of vitamin K, but the compound binds tightly to thylakoid membranes and tends to have low bioavailability. Vitamin K2 is produced by microbes in the gut and is a primary source of the vitamin. Infants in the first few days before they establish their gut flora and people taking broad spectrum antibiotics may have reduced levels, as a result. Dietary deficiency is rare in the absence of damage to the small bowel. Others at risk of deficiency include people with chronic kidney disease and anyone suffering from a vitamin D deficiency. Deficiencies produce symptoms of easy bruising, heavy menstrual bleeding, anemia, and nosebleeds.

Steroids, such as cholesterol are found in membranes and act as signaling hormones in traveling through the body.

Steroid hormones are all made from cholesterol and are grouped into five categories - mineralocorticoids (21 carbons), glucocorticoids (21 carbons), progestagens (21 carbons), androgens (19 carbons), and estrogens (18 carbons).

Mineralocorticoids

Mineralocorticoids are steroid hormones that influence water and electrolyte balances. Aldosterone (Figure 2.238) is the primary mineralocorticoid hormone, though other steroid hormones (including progesterone) have some functions like it. Aldosterone stimulates kidneys to reabsorb sodium, secrete potassium, and passively reabsorb water. These actions have the effect of increasing blood pressure and blood volume. Mineralocorticoids are produced by the zona glomerulosa of the cortex of the adrenal gland.

Glucocorticoids

Figure 2.238 - Aldosterone - A mineralocorticoid

Glucocorticoids (GCs) bind to glucocorticoid receptors found in almost every vertebrate animal cell and act in a feedback mechanism in the immune system to reduce its activity. GCs are used to treat diseases associated with overactive immune systems. These include allergies, asthma, and autoimmune dis- Figure 2.237 - Steroid numbering scheme Image by Pehr Jacobson eases. Cortisol (Figure 2.239) is an important glucocorticoid with cardiovascular, metabolic, and immunologic functions. The synthetic glucocorticoid known as dexamethasone has medical applications for treating rheumatoid arthritis, bronchospasms (in asthma), and inflammation due to its increased potency (25-fold) compared to cortisol. Glucocorticoids are produced primarily in the zona fasciculata of the adrenal cortex.

Progestagens

Figure 2.239 - Cortisol - A glucocorticoid

Progestagens (also called gestagens) are steroid hormones that work to activate the progesterone receptor upon binding to it. Synthetic progestagens are referred to as progestins. The most common progestagen is progesterone (also called P4 - Figure 2.240) and it has functions in maintaining pregnancy. Progesterone is produced primarily in the diestrus phase of the estrous cycle by the corpus luteum of mammalian ovaries. In pregnancy, the placenta takes over most progesterone production.

Figure 2.240 Progesterone - A progestagen

Androgens are steroid hormones that act by binding androgen receptors to stimulate development and maintenance of male characteristics in vertebrates. Androgens are precursors of estrogens (see below). The primary androgen is testosterone (Figure 2.241). Other important androgens include dihydrotestosterone (stimulates differentiation of penis, scrotum, and prostate in embryo) and androstenedione (common precursor of male and female hormones).

Figure 2.241 - Testosterone - An androgen

The estrogen steroid hormones are a class of compounds with important roles in menstrual and estrous cycles. They are the most important female sex hormones. Estrogens act by activating estrogen receptors inside of cells. These receptors, in turn, affect expression of many genes. The major estrogens in women include estrone (E1), estradiol (E2 - Figure 2.242), and estriol (E3). In the reproductive years, estradiol predominates. During pregnancy, estriol predominates and during menopause, estrone is the major estrogen.

Figure 2.242 - Estradiol - An estrogen

Estrogens are made from the androgen hormones testosterone and androstenedione in a reaction catalyzed by the enzyme known as aromatase. Inhibition of this enzyme with aromatase inhibitors, such as exemestane, is a strategy for stopping estrogen production. This may be part of a chemotherapeutic treatment when estrogenresponsive tumors are present.

Cannabinoids

Cannabinoids are a group of chemicals that bind to and have effects on brain receptors (cannabinoid receptors), repressing neurotransmitter release. Classes of these compounds include endocannabinoids (made in the body), phytocannabinoids (made in plants, such as marijuana), and synthetic cannabinoids (man-made).

Endocannabinoids are natural molecules derived from arachidonic acid. Cannabinoid receptors are very abundant, comprising the largest number of G-protein- 247 Figure 2.243 - Tetrahydrocannabinol - Active ingredient in marijuana coupled receptors found in brain. The best known phytocannabinoid is &Delta-9- tetrahydrocannabinol (THC), the primary psychoactive ingredient (of the 85 cannabinoids) of marijuana (Figure 2.243).

Figure 2.243 - Tetrahydrocannabinol - Active ingredient in marijuana

Figure 2.244 - Anandamide - An endocannabinoid

Anandamide (N-arachidonoylethanolamine - Figure 2.244) is an endocannabinoid neurotransmitter derived from arachidonic acid. It exerts its actions primarily through the CB1 and CB2 cannabinoid receptors, the same ones bound by the active ingredient in marijuana, &Delta9-tetrahydrocannabinol. Anandamide has roles in stimulating eating/appetite and affecting motivation and pleasure. It has been proposed to play a role in &ldquorunners high,&rdquo an analgesic effect experienced from exertion, especially among runners. Anandamide appears to impair memory function in rats.

Anandamide has been found in chocolate and two compounds that mimic its effects (N-oleoylethanolamine and Nlinoleoylethanolamine) are present as well. The enzyme fatty acid amide hydrolase (FAAH) breaks down anandamide into free arachidonic acid and ethanolamine.

Lipoxins (Figure 2.245) are eicosanoid compounds involved in modulating immune responses and they have anti-inflammatory effects. When lipoxins appear in inflammation it begins the end of the process. Lipoxins act to attract macrophages to apoptotic cells at the site of inflammation and they are engulfed. Lipoxins further act to start the resolution phase of the inflammation process.

Figure 2.245 - Lipoxin A4

At least one lipoxin (aspirin-triggered LX4) has its synthesis stimulated by aspirin. This occurs as a byproduct of aspirin&rsquos acetylation of COX-2. When this occurs, the enzyme&rsquos catalytic activity is re-directed to synthesis of 15R-hydroxyeicosatetraenoic acid (HETE) instead of prostaglandins. 15R-HETE is a procursor of 15-epimer lipoxins, including aspirin-triggered LX4.

Figure 2.246 - Structure of heme B

Heme groups are a collection of protein/ enzyme cofactors containing a large heterocyclic aromatic ring known as a porphyrin ring with a ferrous (Fe++) ion in the middle. An example porphyrin ring with an iron (found in Heme B of hemoglobin), is shown in Figure 2.246. When contained in a protein, these are known collectively as hemoproteins (Figure 2.247).

Heme, of course, is a primary component of hemoglobin, but it is also found in other proteins, such as myoglobin, cytochromes, and the enzymes catalase and succinate dehydrogenase. Hemoproteins function in oxygen transport, catalysis, and electron transport. Heme is synthesized in the liver and bone marrow in a pathway that is conserved across a wide range of biology.

Figure 2.247 - Heme embedded in the succinate dehydrogenase hemoprotein Wikipedia

Porphobilinogen

Porphobilinogen (Figure 2.248) is a pyrrole molecule involved in porphyrin metabolism. It is produced from aminolevulinate by action of the enzyme known as ALA dehydratase. Porphobilinogen is acted upon by the enzyme porphobilinogen deaminase. Deficiency of the latter enzyme (and others in porphyrin metabolism) can result in a condition known as porphyria, which results in accumulation of porphobilinogen in the cytoplasm of cells.

Figure 2.248 - Porphobilinogen

The disease can manifest itself with acute abdominal pain and numerous psychiatric issues. Both Vincent van Gogh and King ` George III are suspected to have suffered from porphyria, perhaps causing the &ldquomadness of King George III.&rdquo Porphyria is also considered by some to be the impetus for the legend of vampires seeking blood from victims, since the color of the skin in non-acute forms of the disease can be miscolored, leading some to perceive that as a deficiency of hemoglobin and hence the &ldquothirst&rdquo for blood imagined for vampires.

Figure 2.249 - Structure of dolichol pyrophosphate

Dolichol is a name for a group of non-polar molecules made by combining isoprene units together. Phosphorylated forms of dolichols play central roles in the N-glycosylation of proteins. This process, which occurs in the endoplasmic reticulum of eukaryotic cells, begins with a membrane-embedded dolichol pyrophosphate (Figure 2.249) to which an oligosaccharide is attached (also see HERE). This oligosaccharide contains three molecules of glucose, nine molecules of mannose and two molecules of N-acetylglucosamine.

Interestingly, the sugars are attached to the dolichol pyrophosphate with the pyrophosphate pointing outwards (away from) the endoplasmic reticulum, but after attachment, the dolichol complex flips so that the sugar portion is situated on the inside of the endoplasmic reticulum. There, the entire sugar complex is transferred to the amide of an asparagine side chain of a target protein.

The only asparagine side chains to which the attachment can be made are in proteins where the sequences Asn-X-Ser or Asn-X-Thr occur. Sugars can be removed/added after the transfer to the protein. Levels of dolichol in the human brain increase with age, but in neurodegenerative diseases, they decrease.

Figure 2.250 - Pine tree resin - A source of terpenes Wikipedia

Terpenes are members of a class of nonpolar molecules made from isoprene units. Terpenes are produced primarily by plants and by some insects. Terpenoids are a related group of molecules that contain functional groups lacking in terpenes.

Terpenes have a variety of functions. In plants, they often play a defensive role protecting from insects. The name of terpene comes from turpentine, which has an odor like some of the terpenes. Terpenes are common components of plant resins (think pine) and they are widely used in medicines and as fragrances. Hops, for example, gain some of their distinctive aroma and flavor from terpenes. Not all terpenes, however have significant odor.

Figure 2.251 - Three monoterpenes

Terpenes, like steroids, are synthesized starting with simple building blocks known as isoprenes. There are two of them - dimethylallyl pyrophosphate and the related isopentenyl pyrophosphate and (Figures 2.252 and 2.253) which combine 1-2 units at a time to make higher order structures. Terpene synthesis overlaps and includes steroid synthesis.

Figure 2.252 - Dimethylallyl pyrophosphate

Terpenes and terpenoids are classified according to how many isoprene units they contain. They include hemiterpenes (one unit), monoterpenes (two units), sesquiterpenes (three units), diterpenes (four units), sesterterpenes (five units), triterpenes (six units), sesquarterpenes (seven units), tetraterpenes (eight units), polyterpenes (many units). Another class of terpene-containing molecules, the norisoterpenoids arise from peroxidase-catalyzed reactions on terpene molecules.

Figure 2.253 - Isopentenyl pyrophosphate

Common terpenes include monoterpenes of terpineol (lilacs), limonene (citrus), myrcene (hops), linalool (lavender), and pinene (pine). Higher order terpenes include taxadiene (diterpene precursor of taxol), lycopene (tetraterpenes), carotenes (tetraterpenes), and natural rubber (polyterpenes).

Steroid precursors geranyl pyrophosphate (monoterpene derivative), farnesyl pyrophosphate (sesquiterpene derivative), and squalene (triterpene) are all terpenes or derivatives of them. Vitamin A and phytol are derived from diterpenes.

Figure 2.254 - Lycopene - A tetraterpene

Caffeine is the world&rsquos most actively consumed psychoactive drug (Figure 2.255). A methylxanthine alkaloid, caffeine is closely related to adenine and guanine and this is responsible for many effects on the body. Caffeine blocks the binding of adenosine on its receptor and consequently prevents the onset of drowsiness induced by adenosine. Caffeine readily crosses the blood-brain barrier and stimulates release of neurotransmitters. Caffeine stimulates portions of the autonomic nervous system and inhibits the activity of phosphodiesterase. The latter has the result of raising cAMP levels in cells, which activates protein kinase A and activates glycogen breakdown, inhibits TNF-&alpha and leukotriene synthesis, which results in reduction of inflammation and innate immunity.

Figure 2.255 - Caffeine

Caffeine also has effects on the cholinergic system (acetylcholinesterase inhibitor), is an inositol triphosphate receptor 1 antagonist, and is a voltage independent activator of ryanodin receptors (a group of calcium channels found in skeletal muscle, smooth muscle, and heart muscle cells).

The half-life of caffeine in the body varies considerably. In healthy adults, it has a half-life of about 3-7 hours. Nicotine decreases the half-life and contraceptives and pregnancy can double it. The liver metabolizes caffeine, so the health of the liver is a factor in the halflife. CYP1A2 of the cytochrome P450 oxidase enzyme is primarily responsible. Caffeine is a natural pesticide in plants, paralyzing predator bugs.

Lipoprotein complexes and lipid movement in the body

Lipoprotein complexes are combinations of apolipoproteins and lipids bound to them that solubilize fats and other non-polar molecules, such as cholesterol, so they can travel in the bloodstream between various tissues of the body. The apolipoproteins provide the emulsification necessary for this. Lipoprotein complexes are formed in tiny &ldquoballs&rdquo with the water soluble apolipoproteins on the outside and non-polar lipids, such as fats, cholesteryl esters, and fat soluble vitamins on the inside.

They are categorized by their densities. These include (from highest density to the lowest) high density lipoproteins (HDLs), Low Density Lipoproteins (LDLs), Intermediate Density Lipoproteins (IDLs), Very Low Density Lipoproteins (VLDLs) and the chylomicrons. These particles are synthesized in the liver and small intestines.

Apolipoproteins

Figure 2.256 - Apolipoproteins

Each lipoprotein complex contain a characteristic set of apolipoproteins, as shown in Figure 2.256. ApoC-II and ApoC-III are notable for their presence in all the lipoprotein complexes and the roles they play in activating (ApoC-II) or inactivating (ApoC-III) lipoprotein lipase. Lipoprotein lipase is a cellular enzyme that catalyzes the breakdown of fat from the complexes. ApoE (see below) is useful for helping the predict the likelihood of the occurrence of Alzheimer's disease in a patient.

Gene editing

Figure 2.257 - ApoA-I

ApoB-48 and ApoB-100 are interesting in being coded by the same gene, but a unique mRNA sequence editing event occurs that converts one into the other. ApoB-100 is made in the liver, but ApoB-48 is made in the small intestine. The small intestine contains an enzyme that deaminates the cytidine at nucleotide #2153 of the common mRNA. This changes it to a uridine and changes the codon it is in from CAA (codes for glutamine) to UAA (stop codon). The liver does not contain this enzyme and does not make the change in the mRNA. Consequently, a shorter protein is synthesized in the intestine (ApoB-48) than the one that is made in the liver (ApoB-100) using the same gene sequence in the DNA.

The movement of fats in the body is important because they are not stored in all cells. Only specialized cells called adipocytes store fat. There are three relevant pathways in the body for moving lipids. As described below, they are 1) the exogenous pathway 2) the endogenous pathway, and 3) the reverse transport pathway.

Exogenous pathway

Figure 2.258 - Schematic diagram of a chylomicron Image by Aleia Kim

Dietary fat entering the body from the intestinal system must be transported, as appropriate, to places needing it or storing it. This is the function of the exogenous pathway of lipid movement in the body. All dietary lipids (fats, cholesterol, fat soluble vitamins, and other lipids) are moved by it. In the case of dietary fat, it begins its journey after ingestion first by being solubilized by bile acids in the intestinal tract. After passing through the stomach, pancreatic lipases clip two fatty acids from the fat, leaving a monoacyl glycerol. The fatty acids and monoacyl glycerol are absorbed by intestinal cells (enterocytes) and reassembled back into a fat, and then this is mixed with phospholipids, cholesterol esters, and apolipoprotein B-48 and processed to form chylomicrons (Figures 2.258 & 2.259) in the Golgi apparatus and smooth endoplasmic reticulum.

Figure 2.259 - Another perspective of a chylomicron WIkipedia

These are exocytosed from the cell into lymph capillaries called lacteals. The chylomicrons pass through the lacteals and enter the bloodstream via the left subclavian vein. Within the bloodstream, lipoprotein lipase breaks down the fats causing the chylomicron to shrink and become what is known as a chylomicron remnant. It retains its cholesterol and other lipid molecules.

The chylomicron remnants travel to the liver where they are absorbed (Figure 2.260). This is accomplished by receptors in the liver that recognize and bind to the ApoE of the chylomicrons. The bound complexes are then internalized by endocytosis, degraded in the lysosomes, and the cholesterol is disbursed in liver cells.

Endogenous pathway

The liver plays a central role in managing the body&rsquos needs for lipids. When lipids are needed by the body or when the capacity of the liver to contain more lipids than is supplied by the diet, the liver packages up fats and cholesteryl esters into Very Low Density Lipoprotein (VLDL) complexes and exports them via the endogenous pathway. VLDL complexes contain ApoB-100, ApoC-I, ApoC-II, ApoC-III, and ApoE apolipoproteins. VLDLs enter the blood and travel to muscles and adipose tissue where lipoprotein lipase is activated by ApoC-II. In the muscle cells, the released fatty acids are taken up and oxidized. By contrast, in the adipoctyes, the fatty acids are taken up and reassembled back into triacylglycerides (fats) and stored in fat droplets. Removal of fat from the VLDLs causes them to shrink, first to Intermediate Density Lipoprotein (IDL) complexes (also called VLDL remnants) and then to Low Density Lipoprotein (LDL) complexes.

Figure 2.260 - Movement of lipids in the body - Green = exogenous pathway Blue = endogenous pathway Purple = reverse transport pathway Image by Aleia Kim

Shrinking of VLDLs is accompanied by loss of apolipoproteins so that LDLs are comprised primarily of ApoB-100. This lipoprotein complex is important because cells have receptors for it to bind and internalize it by receptor-mediated endocytosis (Figure 2.261). Up until this point, cholesterol and cholesteryl esters have traveled in chylomicrons, VLDLs, and IDLs as fat has been stripped stripped away. For cholesterol compounds to get into the cell from the lipoprotein complexes, they must be internalized by cells and that is the job of receptormediated endocytosis.

Reverse transport pathway

Another important consideration of the movement of lipids in the body is the reverse transport pathway (Figure 2.260). It is also called the reverse cholesterol transport pathway, since cholesterol is the primary molecule involved. This pathway involves the last class of lipoprotein complexes known as the High Density Lipoproteins (HDLs). In contrast to the LDLs, which are commonly referred to as &ldquobad cholesterol&rdquo (see below also), the HDLs are known as &ldquogood cholesterol.&rdquo

Figure 2.261 - The process of receptor-mediated endocytosis Image by Aleia Kim

HDLs are synthesized in the liver and small intestine. They contain little or no lipid when made (called depleted HDLs), but serve the role of &ldquoscavenger&rdquo for cholesterol in the blood and from remnants of other (damaged) lipoprotein complexes in the blood. To perform its task, HDLs carry the enzyme known as lecithincholesterol acyl transferase (LCAT), which they use to form cholesteryl esters using fatty acids from lecithin (phosphatidylcholine) and then they internalize them.

The cholesterol used for this purpose comes from the bloodstream, from macrophages, and from foam cells (macrophage-LDL complexes - Figure 2.262). Addition of cholesteryl esters causes the HDL to swell and Figure 2.261 - The process of receptor-mediated endocytosis Image by Aleia Kim when it is mature, it returns its load of cholesterol back to the liver or, alternatively, to LDL molecules for endocytosis. HDLs have the effect of lowering levels of cholesterol and it is for that reason they are described as &ldquogood cholesterol.&rdquo

Regulation of lipid transfer

Figure 2.262 - Foam cell aggregate Wikipedia

It is important that cells get food when they need it so some control of the movement of nutrients is critical. The liver, which plays the central role in modulating blood glucose levels, is also important for performing the same role for lipids. It accomplishes this task the use of specialized LDL receptors on its surface. Liver LDL receptors bind LDLs that were not taken up by other cells in their path through the bloodstream. High levels of LDLs are a signal to the liver to reduce the creation of VLDLs for release.

People with the genetic disease known as familial hypercholesterolemia, which manifests with dangerously high levels of LDLs, lack properly functioning LDL receptors on their liver cells.Figure 2.263 - Progression of atherosclerosis Wikipedia Figure 2.263 - Progression of atherosclerosis Wikipedia Figure 2.263 - Progression of atherosclerosis Wikipedia

Figure 2.263 - Progression of atherosclerosis Wikipedia

In sufferers of this disease, the liver never gets the signal that the LDL levels are high. In fact, to the liver, it appears that all VLDLs and LDLs are being taken up by peripheral tissues, so it creates more VLDLs to attempt to boost levels. Untreated, the disease used to be fatal early, but newer drugs like the statins have significantly increased life spans of patients. Cellular needs for the contents of LDLs are directly linked to the levels of synthesis of LDL receptors on their membranes. As cells are needing more cholesterol, their synthesis of components for receptors goes up and it decreases as need diminishes.

Good cholesterol / bad cholesterol

It is commonly accepted that &ldquohigh cholesterol&rdquo levels are not healthy. This is due, at least indirectly, to the primary carriers of cholesterol, the LDLs. A primary function of the LDLs is to deliver cholesterol and other lipids directly into cells by receptor mediated endocytosis (Figure 2.237). High levels of LDLs, though, are correlated with formation of atherosclerotic plaques (Figure 2.263 & 2.264) and incidence of atherosclerosis, leading to the description of them as &ldquobad cholesterol.&rdquo This is because when LDL levels are very high, plaque formation begins. It is thought that reactive oxygen species (higher in the blood of smokers) causes partial oxidation of fatty acid groups in the LDLs. When levels are high, they tend to accumulate in the extracellular matrix of the epithelial cells on the inside of the arteries. Macrophages of the immune system take up the damaged LDLs (including the cholesterol).

Since macrophages can&rsquot control the amount of cholesterol they take up, cholesterol begins to accumulate in them and they take on appearance that leads to their being described as &ldquofoam cells.&rdquo With too much cholesterol, the foam cells, however, are doomed to die by the process of programmed cell death (apoptosis). Accumulation of these, along with scar tissue from inflammation result in formation of a plaque. Plaques can grow and block the flow of blood or pieces of them can break loose and plug smaller openings in the blood supply, ultimately leading to heart attack or stroke.

Good cholesterol

On the other hand, high levels of HDL are inversely correlated with atherosclerosis and arterial disease. Depleted HDLs are able to remove cholesterol from foam cells. This occurs as a result of contact between the ApoA-I protein of the HDL and a transport protein on the foam cell (ABC-G1). Another transport protein in the foam cell, ABCA-1 transports extra cholesterol from inside the cell to the plasma membrane where it is taken up into the HDL and returned to the liver or to LDLs by the reverse transport cholesterol pathway.

Figure 2.264 - Actual carotid artery plaque Wikipedia

Deficiency of the ABCA-1 gene leads to Tangier disease. In this condition, HDLs are almost totally absent because they remain empty as a result of not being able to take up cholesterol from foam cells, so they are destroyed by the body.

ApoE and Alzheimer&rsquos disease

ApoE is a component of the chylomicrons and is also found in brain, macrophages, kidneys, and the spleen. In humans, it is found in three different alleles, E2, E3, and E4. The E4 allele (present at about 14% of the population) is associated with increased likelihood of contracting Alzheimer's disease. People heterozygous for the allele are 3 times as likely to contract the disease and those homozygous for it are 15 times as likely to do so. It is not known why this gene or allele is linked to the disease. The three alleles differ only slightly in amino acid sequence, but the changes do cause notable structural differences. The E4 allele is associated with increased calcium ion levels and apoptosis after injury. Alzheimer&rsquos disease is associated with accumulation of aggregates of the &beta- amyloid peptide. ApoE does enhance the proteolytic breakdown of it and the E4 isoform is not as efficient in these reactions as the other isoforms.