SUBJECT: BIOCHEMISTRY TOPIC: LIPID METABOLISM 1 (Chapter 22

:
Oxidation of Fatty Acids: Ketogenesis & Chapter 23: Biosynthesis of Fatty acids and Eicosanoids )

LECTURER: Dr. LAYGO DATE: 11/23/2010

There is a relationship between intake of carbohydrates and lipid synthesis. Predominating hormone during sembreak (eat, sleep, rest) is insulin. Glucose derived from carbohydrates undergoes glycolysis and is stimulated in the presence of insulin. There are enzymes that are induced* with the presence of insulin. Insulin also stimulates fatty acid synthesis. That is the time when our cells have the luxury of synthesizing fatty acids and there by storing them as triacylglycerol. Triacylglycerol is a molecule with glycerol backbones, and in the carbons of this glycerol are esterified fatty acids.

If the cells need energy, pyruvate will be brought inside the mitochondrial matrix and be oxidatively decarboxylated to form Acetyl CoA. β- oxidation o Process whereby cells try to degrade fatty acids o Beta because beta carbon (or carbon number 3) is the one involved  Beta carbon is transformed into a carbonyl carbon (carbon attached to oxygen with a double bond)  First 3 steps utilized by the cells in order to liberate 2 carbon moieties from fatty acids  For cells to degrade fatty acids, 2 carbons should be removed at a time General Types of pathways: o Linear pathway  Glycolysis  Glucose  Pyruvate (or lactate in a different case) o Cyclic pathway  Kreb’s cycle  Every turn gives rise to OAA o Spiral pathway  Fatty acid synthesis  β- oxidation of fatty acid

* Induced/ induction involves 2 processes: transcription and translation. Wherein in enzymes that are critical for stimulation of glycolysis will be synthesized in more amounts such as Fructose 1,6-Bisphosphate and Pyruvate kinase. ** Fatty acid #1 is usually palmitate, Fatty acid #2 is usually oleate, Fatty acid #3 is oleate or a polyunsaturated fatty acyl group.

Every time a cell removes 2 carbon at a time, the cell utilizes 4 reactions.  Synthesis of fatty acids is the exact opposite of how the cells break them down.

I. FATTY ACID SYNTHESIS:  Needs activated intermediates that are bound to a particular molecule, example of the molecule of which activate intermediates are bound is: o ACP – Acyl Carrier Protein Needs a source of electrons and protons o NADPH + H coming from hexose monophosphate (alternative carbohydrate metabolic pathway) pathway and can also come from citrate shuttle system (aka Acetyl-CoA shuttle system) Found in the cytoplasm, enzymes that will carry the different steps are coming from the cytosol. To carry on the elongation of carbons to make up the fatty acid, specifically the palmitic acid, we need an enzyme complex known as Fatty Acid Synthase Enzyme Complex. After intake a lot of carbohydrates, this will go through glycolysis producing the pyruvate.

Franz Knoop elucidated the different steps initially making up the β- oxidation or the breakdown of fatty acids. And since the breakdown of fatty acids is in spiral pathway, he hypothesized that synthesis of these is exactly in the opposite way. And later on he was proven right. Requirements of Fatty Acid Synthesis:  Acetyl CoA o Carbohydrates in the form of glucose after being transformed to pyruvate enters the mitochondrial matrix to be oxidatively decarboxylated to form Acetyl CoA o The one initially utilized to synthesize fatty acids o Acted upon by an enzyme called Acetyl CoA Carboxylase or ACC enzyme  Catalyzes the carboxylation reaction producing Malonyl CoA

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Catalyzes the rate limiting step for synthesis fatty acids  Single, multifunctional enzyme  Requires a biotin group which is attached to a lysine residue of a protein molecule  Contains an allosteric regulatory site  Citrate – most important regulatory factor  Catalyzes the rate-limiting step for fatty acid synthesis which forms malonyl CoA  Long chain fatty acyl CoAse  Inhibits Acetyl CoA Carboxylase o Initial building block o 2-carbon moiety o 16th and 15th carbon of palmitic acid (most common fatty acid synthesized by the cell) came from Acetyl CoA and the rest came from malonyl CoA CO2 in the form of bicarbonate o Source of carbon ATP o Source of energy to form a covalent bond Biotin from biocytin o Being a carboxylation reaction wherein the the source of carbon is CO2, it is the coenzyme needed by Acetyl CoA Carboxylase Malonyl CoA o Utilized by the cell to lengthen the carbons from Acetyl CoA o As a result of carboxylation reaction, it is 3-carbon moiety o It is the source of the 2 carbons used to elongate Acetyl CoA o In utilizing a 3-carbon moiety, remember that the cells only need 2 carbons, so the other one is liberated as CO2 

o

We start of Acetyl CoA and we elongate that by utilizing 4 different reactions utilized by the cell every cycle (every time a cell adds 2 Carbon Moiety to Acetyl CoA):  Condensation  1st Reduction  Dehydration  2nd Reduction

Answer: o 7 cycles or spirals needed  We have an original 2 carbon moiety from Acetyl CoA and cells try to elongate this by adding 2 carbons at a time (1 spiral/cycle). So initially 2 carbon moiety plus 2 carbons added per cycle (7 cycles) is equal to 16 carbons which is the number of carbon of palmitic acid. 2 + (2 x 7) = 16

Question: How many NADPH + H will the cells need to be able to synthesize a 16-carbon saturated fatty acid?  Answer: If the cell utilizes 7 cycles, and each of this cycle involves 2 reduction reactions, it needs 14 NADPH + H because each reduction reaction needs one NADPH + H. For every cycle, we need 2 NADPH+H because we have 2 reduction reactions

Malonyl CoA   Has 3 carbons Source of 2 carbons used to elongate Acetyl CoA o We only need 2 carbons, and 1 carbon should be eliminated in the form of Carbon Dioxide. Is taken over by fatty acid synthase enzyme complex, thereby elongating acetyl CoA to eventually form palmitic acid. Signal for the cell to synthesize fatty acids o It’s a negative allosteric effector for fatty acid oxidation  Cells are endowed with a lot of energy and therefore can synthesize a lot of fatty acids

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Question: If the cells will be synthesizing a 16-carbon fatty acid, how many spirals or cycles will the cells need in order to form palmitic acid (most common fatty acid synthesized by the cell)?

 
Structure of Palmitic acid

Remember: o Starting material is a 2-carbon moiety, Acetyl CoA  16th and 15th carbon of Palmitic acid came from the initial Acetyl CoA  The rest came from Malonyl CoA o Acetyl CoA Carboxylase (ACC) is only initially needed to synthesize Malonyl CoA  After the cells have acted upon the initial Acetyl CoA by means of Acetyl CoA Carboxylase, forming Malonyl CoA. Then the fatty acid synthase enzyme complex will now take over the process of elongation. o Fatty acid synthesis is a spiral pathway.

Fatty acid Synthase complex  A dimer o Arranged in a head-to-tail manner

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A large molecule Multiple domain

3-OHACYL dehydrase

Alpha beta transbutenoyl ACP Butyryl ACP

Dehydration

Enoyl Reductase

2nd Reduction

STOICHIOMETRY OF PALMITATE SYNTHESIS: 8ACETYLCoA +7ATP + 14NADPH  PALMITATE + 14NADP++ 8CoASH +6H2O + 7ADP + 7Pi
We need 8 Acetyl CoA molecules, and then from it, we synthesize 7 Malonyl CoA), 7 ATP, 14 NADPH+H forming palmitate, releasing 14 oxidized form of NADP, CoA, water, and 7 ADP and 7 inogrganic phosphates. It being a dimmer, they are arranged into a head to tail manner. These subunits carry on the different 4 reactions that are utilized by the cell every cycle of the pathway.  Transacylation reaction  Initial reaction that attaches and activates intermediates for fatty acid synthesis to ACP (Acyl Carrier Protein).  Acetyl transacylase o Enzyme that catalyzes the binding of acetyl CoA to the ACP  Malonyl transacylase o Enzyme that catalyzes the binding of malonyl CoA to the ACP  Coenzyme A o Counterpart of Fatty Acid Synthase in Beta-oxidation CITRATE  During starvation (glucagon and epinephrine are predominating hormones), if the cells need more energy, Acetyl CoA will be shuttled to kreb’s cycle, and will be condensed with OAA, catalyzed by citrate synthase to form citrate. If the cells are endowed with a lot of energy after a lot of carbohydrate intake, we will be bombarding the citric acid cycle with a lot of citrate which cannot be accommodated by the next step (aconitase and isocitrate dehydrogenase step), so excess of citrate will go out into the cytoplasm (Citrate Shuttle System)

CITRATE SHUTTLE SYSTEM/ACETYL CoA SHUTTLE SYSTEM:

2 reductions and therefore require NADPH+ Needs 7 cycles, 7 malonyl therefore, 14 NADPH+H Enzymes that catalyzes the 4 different reactions: Enzyme Product Reaction catalyzed
Condensation

3-Ketoacyl ACP synthase 3-Ketoacyl ACP reductase

Acetoacetyl ACP

3-OH Butyryl ACP

1st Reduction

Inner mitochondrial membrane is highly selective. It would not allow just the entry and exit of molecules. In the cytoplasm, there will be occurrence of glycolysis to form a lot of pyruvate. Pyruvate goes into the matrix through pyruvate translocase situated in the inner membrane. Pyruvate in the mitochondrial matrix will then be oxidatively decarboxylated by pyruvate dehydrogenase (enzyme complex) to become acetyl CoA. Acetyl CoA will condense with oxaloacetate to form citrate with the help of citrate synthase. If there is a lot of energy, excess of citrate will go out of the cytoplasm and will be converted back to acetyl-

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coA and oxaloacetate with the help of ATP dependent citrate lyase enzyme. Acetyl CoA will be acted upon by Acetyl CoA carboxylase to form malonyl-CoA and thereby will be taken over by fatty acid synthase to form fatty acids (such as palmitate). Oxaloacetate will be subsequently reduced by malate dehydrogenase cytosolic with the help of NADH to form malate. Malate can either go again inside the mitochondrial matrix or it can be oxidatively decarboxylated by malate enzyme (or NADP-dependent malate dehydrogenase enzyme) with the help of NADP, which after oxidizing malate back to pyruvate, the cells generate NADPH+H (much needed reducing equivalent needed for fatty acid synthesis). Therefore, if we will try to count the number of citrate molecules that goes out into the cytoplasm, creating acetyl CoA molecules that the cells utilize for fatty acid synthesis, How many of this will be generated, that will contribute to the 14 NADPH+H to complete the synthesis of palmitic acid ?     NADPH+H = 14 Acetyl CoA needed by the cell to synthesize palmitic acid = 8 Citrate = 8 (because from 8 citrates, 8 acetyl CoA will be formed by citrate lyase enzyme) OAA= 8 converted to pyruvate  (8 NADPH+H is generated)

structures. For them to be able to transmit their message inside the cell, they must possess their receptors expressed on the membrane. Epinephrine (amine hormone) is derived from tyrosine. The receptor in the plasma membrane, epinephrine and glucagon have to bind to the receptor, forming a hormone-receptor complex to activate the G-protein. Activated G-protein then activates AC (Adenylyl cyclase), that tries to cyclycize ATP to form the 2nd messenger, which is cAMP (Cyclic AMP). cAMP activates the inactive protein kinase A. If there is an activated protein kinase A, proteins and enzymes will receive phosphate groups. Therefore, these enzymes and proteins are inactivated due to their phosphorylation. There is a predominance of epinephrine and glucagon during starvation. Insulin is exerting the opposite effect of glucagon. Insulin is also a peptide hormone, which has a receptor on the membrane. The receptor of insulin is called receptor tyrosine kinase. So from this moment, when it binds, after forming a hormone-receptor complex, one of the actions is to stimulate an enzyme called phosphodiestarase (3’,5’-cyclic AMP phosphodiesterase) to transform cAMP to 5’-AMP. Therefore, protein kinase is not activated… Enzymes will be in the dephosphorylated form. Activated form of Acetyl CoA carboxylase is in dephosphorylated form, therefore can do its job to transform Acetyl CoA into Malonyl CoA. But with the predominance of glucagon and epinephrine, Acetyl CoA is inhibited. Remember: Enzymes catalyze a specific reactions and therefore specific names:
Protein kinase A  activation is dependent on cAMP. Protein kinase G  activation is dependent cGMP (a second messenger that activates protein kinase G) Protein kinase C  activation is dependent on calcium ions (C2+)

***14 NADPH+H – 8 NADPH+H = remaining 6 NADPH+H (which can be provided by HMP or Hexose Monophosphate Pathway) As long as we have citrate in the cytoplasm, Acetyl CoA Carboxylase is stimulated. Inactive form Acetyl CoA carboxylase is in the form of monomers. If there are 40 monomers, they try to aggregate to form the active form (protein-protein interaction).

Covalent modification:   Attachment of phosphate group to enzymes and proteins One way of which the cells try to stimulate/ inhibit 2 opposing pathways o Ex: Glycogenolysis & Glycogenesis (Carbohydrate metabolism) By attaching a phosphate through glucagon Glucagon and epinephrine  1st messengers (they can transmit messages inside the cells through cAMP dependent protein kinase A cascade). Through covalent modifications whereby activity of most enzymes are activated, while some are inhibited. o So.. Depende sa enzymes.. Merong ibang enzymes na pag naka-tanggap ng phosphate group, ma-aactivate, meron naman iba na ma-iinhibit.  Sa ACC, gumagana lang siya kapag dephosphorylated sila. So dapat walang phosphate group. o Our cells do not have the luxury to synthesize molecules when there’s a need for energy. When there is a need for energy, cells mostly induce catabolism to release a lot of electrons and protons to be harnessed in ETC. After being carried by coenzymes such as FAD and NADH+H. If we transfer electrons and protons through NADH+H, it will create a PO ratio

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There is insulin predominance during FA synthesis. The confirmation of Acetyl CoA carboxylase is in the dephosphorylated form (activated form). Insulin and glucagon are peptide hormones, so they cannot just traverse the plasma membrane due to the nature of the

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of 3:1 (3 moles of ATP per 1 mole of NADH + H) and ½ of oxygen molecule (O2) or 1 oxygen atom will be used as electron acceptor to form metabolic H2O. If FADH2 (2:1), 2 moles of ATP as against 1 atom of oxygen that serves as the final electron acceptor to form metabolic water. The opposite is brought about by the predominance of insulin  removal of phosphate group o Wherein it stimulates phosphodiesterase not adenylyl cyclase o Insulin also stimulates phosphatase  Phosphatase removes phosphate group.

and series 3 leukotrienes). Furthermore, through a desaturase delta 5, there will be an introduction of double bonds in between carbons 5 and 6 to form arachidonic acid (precursor of series 2 prostaglandins and series 4 leukotrienes). This can also be elongated after elongation desaturation to form another precursor for eicosanoid synthesis called tinodonic acid or eicosapentaenoic acid (precursor of series 3 prostaglandins or series 5 leukotrienes)

dihomo-gamma-linolenic acid 

Therefore, this will now effect attachment of a phosphate, enzymes in the cytoplasm when interact with protein kinase A, are activated while some are inhibited.

Introduce a double bond, and elongate the FA to form a C18, C20, C22 FA. For introduction of double bond in a saturated fatty acid, we need a source of electrons and protons coming form NADH+H with the help of cytochrome B5 reductase, in the participation of FAD. NADH+H transfers electrons to FAD become reduced to FADH2, that again will be oxidized. FADH2 will transfer electrons and protons to the ferric form (Fe3+) that will be reduced to ferrous (Fe2+) forming a double bond, with water molecule removal. After introduction of double bond, elongation process is started. Example: LINOLEIC ACID: 18 carbon with 2 double bonds situated in carbon 9 and 10, and carbons 12 and 13. Initially, there will be an introduction of another double bond between carbons 6 and 7.
Remember: Polyunsaturated fatty acids have 2 or more double bonds, but after desaturase activity, it will become 3, 6, 9, 12.

Concept of elongation: Introducing a double bond (action of desaturase) and adding a carbon atom (elongase) alternately. So initially desaturation, elongation, desaturation, and so on.

SHORT TERM REGULATION OF FATTY ACID SYNTHESIS with the predominance of INSULIN.

Then there will be an elongation process wherein there is addition of 2 carbon moiety through elongase enzyme to form a 20-carbon  dihomo-gamma-linolenic acid or eicosatrienoic acid (precursor of series 1 prostaglandins

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TRANSPORT OF FATTY ACIDS IN THE BLOODSTREAM: 1. PLASMA ALBUMIN a. b. c. Major carrier of free fatty acids in blood Binds ~6 molecules of fatty acids per 1 molecule of albumin Also binds and transports bilirubin, other organic anions, and a variety of xenobiotic compounds (aspirin, barbiturates, Coumadin, oral hypoglycaemic agents)

In β- oxidation, Malonyl CoA inhibits carnitine acyl transferase 1 (enzyme needed for beta oxidation) Insulin  Induces glycolysis, therefore forms a lot of pyruvate, a lot of Acetyl CoA, a lot of citrate which cannot be accommodated by kreb’s cycle and goes out to the cytoplasm and is cleaved into acetyl CoA and OAA. Acetyl CoA will be acted upon by acetyl CoA carboxylase. More citrate will further stimulate allosterically your acetyl CoA carboxylase activity, thereby forming palmitate. When it is activated, and bound to molecule, forms palmitoyl CoA. From malonyl to palmitate, we will see the action of fatty acid synthase complex (FAS).

2. LIPOPROTEINS a. Carriers of esterified fatty acids, primarily as triaglycerols

MOBILIZATION OF STORED TRIAGLYCEROL

Lipoproteins, specifically chylomicrons which carry dietary triacylglycerol, while VLDL carries the endogenously synthesized triacylglycerol into the circulation.

TRANSLOCATION OF FATTY ACIDS INTO MITOCHONDRION

Epinephrine, glucagon  deactive Protein Kinase A will be formed and acetyl CoA carboxylase will be phosphorylated then transforming it into its inactive form. In times of starvation, we need a lot of energy, and there must be mobilization of stored triacylglycerol. In the presence of glucagon and epinephrine, through cAMP dependent protein kinase cascade, there will be phosphorylation of hormone-sensitive lipase (sensitive to glucagon and epinephrine). So now, these hormone sensitive lipase being phosphorylated, thereby activated, will now try to hydrolyze triacylglycerol into fatty acids and eventually into glycerol backbone. Fatty acids will be carried into circulation. And in the circulation, will be carried by albumin protein. 6 molecules of fatty acids in 1 albumin, going back to the liver and is subjected to Beta-oxidation.

Lingual lipase  initial digestion of lipids in the mouth Chyme goes down into duodenum and stimulates the secretion of pancreatic juice. Chyme contains lipids/triacyglycerol, proteins, carbohydrates so there is a variety of enzymes that permits digestion. In this lecture,

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we will focus on the enzyme, PANCREATIC LIPASE since it is the one responsible for lipid digestion. Pancreatic lipase acts on dietary triacylglycerols which permits the release 2 free fatty acids and a 2monoacylglycerol (there is still a fatty acid esterified at 2nd carbon of glycerol backbone). Pancreatic lipase is stabilized by colipase. Before the action of pancreatic lipase, there is an initial step in which the cells have a way to decerase the lipid-water interphase so that the hydrophilic enzymes can interact with hydrophobic triacyglycerols (lipid droplets), which we call emulsification. After that, pancreatic lipase takes effect. Upon the breakdown, the fates of short and medium chain fatty acids will be immediately transported into the circulation, and will undergo oxidation in the mitochondrion of the liver cells. While long chain fatty acids together with 2-monoacylglycerols (after absorption by intestinal cells), they will resynthesize triacylglycerol inside intestinal cell and will be given a coat of apoliprotein B48 to form chylomicrons and to which the chylomicrons transport them to the lymphatic system and will eventually be emptied into the thoracic duct and then to subclavian vein. Fransup  said that FA must be degraded by removal of 2 carbons at a time, which occurs in mitochondrion (by Leninger). And 2 carbons released is not acetate, but acetyl-CoA (FA synthesis is also called as Linen Cycle) BETA-OXIDATION OF FATTY ACIDS  Beta carbon is the one involved in the process of cleavage i. By transforming this 3rd carbon into carbonyl/carboxyl carbon ii. Done by the 1st 3 steps exactly opposite of the Fatty Acid synthesis (Condensation, Reduction, Dehydration) 1. Oxidation 2. Hydration 3. 2nd Oxidation 4. Cleavage Cells liberate phenylacetate  true for even chain fatty acids Cells liberate benzoic acid  true for odd chain fatty acids Cell degrade fatty acids by removal of 2 carbons at a time For fatty acid oxidation: i. Molecule attached to intermediate is attached by Coenzyme A (counterpart of transacylase in fatty acid synthesis) which is carried out by Acetyl CoA Synthetase ii. 2-step reaction 1. ATP is hydrolyzed to AMP + Pyrophosphate 2. Pi is hydrolyzed by pyrophosphatase to liberate 2 Pi a. Energy liberated by 1 mole of Pyrophosphate being hydrolyzed to Pi = -6.6 kCal i. If this is the kind of hydrolysis carried out, it amounts to 2 moles of ATP

ii.

Acyl CoA Synthetase   

Hydrolysis of pyrophosphate to 2 inorganic phosphates ensures the completion of the reaction.

Also known as thiokinase Catalyzes the activation of acyl groups, forming acyl CoA Ex: Palmitic acid + Coenzyme A  Palmitoyl CoA o Can now be allowed to enter the matrix. o Acted upon by Carnitine-Palmitoyl transferase 1

CARNITINE –PALMITOYL TRANSFERASE 1 Allows the transfer of acyl group to carnitine to form acyl carnitine, subsequently releasing coenzyme A. As acyl carnitine, it can now be translocated inside the matrix through the help of a protein embedded in the inner mitochondrial membrane, which is named as carnitine-acyl carnitine translocase. Inside the matrix, this acyl carnitine will be subsequently acted upon by carnitine-palmitoyl transferase 2, transferring acyl group back to Coenzyme A, releasing carnitine. Carnitine will be translocated outside into the intermembranous space, by means of the translocase, in time for the second load of palmitoyl-Coa
**Carnitine-Acyl Transferase 1 is just a general name, while CarnitinePalmitoyl Transferase 1 is a more specific one.

PROTEINS FOR FATTY ACID TRANSPORT

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Site of action of malonyl CoA inhibits carnitine-palmitoyl transferase 1. Carnitine-palmitoyl transferase 1. Malonyl CoA is the rate limiting for beta-oxidation. Carnitine is a molecule is synthesized by liver and kidney cells from Lysine and Methionine. In the biostatic pathway, there are 2 enzymes that need the coenzyme vitamin, ascorbic acid. Different steps repeated every cycle in the pathway of Boxidation:

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the mitochondrion, palmitate must be activated and therefore must be attached to coenzyme A through acyl CoA synthetase. And in the process, it utilizes 2 moles of ATP. Therefore if we would like to know the net amount of ATP produced, we have to subtract 2 moles of ATP to know the net amount of ATP is 129 moles per 1 mole of palmitic acid.
For additional information: In the 1st step: OXIDATION:   1.5~ 2 ATP is the original amount for FADH2 2.5~ 3 ATP is the original amount for NADH+H

ODD CHAIN FATTY ACIDS   Oxidation carried out by Acyl CoA dehydrogenase with the help of FAD (the one that will receive protons and electrons), which is subsequently reduced to FADH2. FAD can easily transfer to ETC, creating a PO ration of 2:1. After the 1st oxidation, the product is trans delta 2 Fatty enoyl CoA. Through hydration, the double bond is hydrated and there is formation of L-beta-Hydroxy acyl CoA with the help of enoyl CoA hydratase and a water molecule. After hydration reaction, comes the second oxidation with the help of NAD with the help carried out by L-hydroxy acyl CoA dehydrogenase, forming NADH+H which goes to electron transport chain, creating a PO ratio of 3:1. Finally, the betacarbon becomes the carbonyl carbon. With the help of Coenzyme A, the sulfur atom in it carries a lone pair of electrons. Oxygen is more electron negative than carbon, so it is a neutrophile that attracts electron to itself, which makes carbon more positive. If we have there a nucleophile, it can easily attack there through a nucleophilic attack, cleaving the bond and releasing acetyl CoA, shortening original FA by 2 carbons and the cycle will go on. If FA is palmitic acid, there is 7 cycles, therefore yields 1 mole of FADH2 and 1 mole of NADH+H per cycle, 5 ATP generated per cycle (because this will create 2(from FADH2 +3 from NADH + H). How many moles of acetyl CoA from Boxidation of palmitic acid? 8 moles of Acetyl CoA. How many moles of ATP? How many FADH2? = 7 How many NADH+H = 7 8 Acetyl CoA goes to the TCA cycle to synthesize more energy during starvation. Remember that 1 mole of Acetyl CoA, when enters the TCA cycle yields 12 ATPs. Gross amount of ATP generated out of complete betaoxidation of palmitic acid is (12ATP x 8) + 14 ATP + 21 ATP = 131 gross ATP per mole of palmitic acid 2 moles of ATP utilized in acyl CoA synthetase to activate FA to form palmitoyl CoA. Palmitic acid that will be oxidized through Beta-oxidation came from the deposited triacylglycerol through the action of HSL (Hormone Sensitive Lipase). Palmitate is carried by albumin in the circulation to bring it back to the liver. Before it enters to Can also be utilized and can be subjected to normal B-oxidation 15-carbon FA  beta-oxidation removes up to the 12th carbon. The last 3 corresponds to carbon number 1, alpha carbon (2nd carbon), and beta carbon (3rd carbon). The resulting 3-carbon moiety in the form of Propionyl CoA can still be utilized by the cell to generate energy.

Propionyl CoA is carboxylated through propionyl CoA to form D-Methylmalonyl CoA and this will be isomerized by epimerase to form L-Methyl Malonyl CoA and through methylmalonyl CoA mutase, forms succinyl CoA. Succinyl CoA is an intermediate of Kreb’s cycle. With this respect, form L-Methylmalonyl CoA to be catalyzed by mutase to form Succinyl CoA. Mutase is one of the 2 enzymes in which vitamin B12 (in the form of 5deoxyadenosyl cobalamine) is needed. While the other enzyme that needs vitamin B12 is methionine synthase enzyme which catalyzes the synthesis of methionine from homocystein. This methionine synthase needs vitamin B12 (in the form of Methyl Cobalamine). UNSATURATED FATTY ACIDS  Cells can also utilize them for energy purposes.

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2 kinds of unsaturated fatty acids: 1. Monounsaturated Fatty Acids

Carbon number 1, Carbon number 2 (alpha carbon), Carbon number 3 (beta carbon) has a methyl substituent; our cells do not have an enzyme to act upon the beta carbon with a methyl substituent. The cells then utilize alpha oxidation, wherein the alpha carbon will be the carbonyl carbon, and the carbon number 1 is liberated as CO2. When carbon 1 is removed, the former alpha carbon becomes the carbon number 1, and the former beta carbon will not have branch. So therefore, this can be subjected to the 4 steps that are repeated every cycle of the pathway. If that is the case, the cleavage will be at the site wherein it releases propionyl CoA (a 3-carbon moiety). Propionyl CoA is transformed to Succinyl CoA and eventually enters Kreb’s cycle. Beta carbon can be subjected to a normal beta oxidation, releasing Acetyl CoA (2-carbon moiety). After that, it releases propionyl CoA again.

 
Initially, there is a double bond situated in carbon 9 and  10. Initially, oleoyl CoA is subjected to 3 cycles of normal  B‐oxidation. After removing 6 carbons, the initial carbon  7 becomes carbon 1, carbon 8 now becomes our alpha  carbon, and carbon 9 becomes the beta carbon. The beta  carbon as we can see is now involved in a double bond,  which our cells will not be able to utilize the carbon with  double bond. So the cells isomerizes  cis‐delta‐3‐ Dodecenoyl CoA forming trans‐delta‐2‐Dodecenoyl CoA,  which can be subsequently hydrated with a mole of  water to form L‐beta‐Hydroxy‐Decanoyl. After this we  can now utilize the normal B‐oxidation cycle to degrade  this kind of fatty acid.

Refsun’s Disease refers to the accumulation of phytanic acid especially in the brain, secondary to enzyme deficiencies. OMEGA OXIDATION  Last carbon or the omega carbon is transformed into a carboxylic carbon. o If this happens, both sides can be subjected to normal B-oxidation

2. Polyunsaturated Fatty Acids 2 double bonds are seen between carbons 9 and 10, 12 and 13 and they undergo 3 cycles of beta-oxidation. Afterwhich, the beta carbon is again involved in a cis confirmation of double bond. By means of an isomerase, it can be transformed into a trans-delta-2-enoyl CoA. Cells use isomerase initially, then dehydrogenase, and lastly reductase so that normal B-oxidation of polyunsaturated fatty acids may be take place. PEROXISOMAL OXIDATION   Flavin-dependent reaction Amount of energy in the form of ATP is less because electrons are not transformed to ETC, but instead transferred to a molecular oxygen, forming hydrogen peroxide Hydrogen peroxide is a harmful compound that is degraded to oxygen and water by the catalase enzyme.

SHORT TERM REGULATION FOR FA METABOLISM:

ALPHA OXIDATION    Oxidizes branched chain fatty acids We derive metabolite intermediate coming from chlorophyll (phytanic acid), a branched chain of fatty acid. May also be a source of energy. 

Glucagon and Epinephrine phosphorylate the acetyl CoA carboxylase, which forms cAMP which then activates protein kinase A and therefore phosphorylates, ACC, inhibiting its activity.

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o

So instead of FA synthesis, the predominant pathway is beta-oxidation, forming a lot of acetyl Coa. Acetyl CoA goes to kreb’s cycle.

Acetone + dehydrogenase

Liver is the site of ketogenesis. But with respect to energy utilization, the liver cannot utlize them as source of energy due to the absence of succinyl CoA -aceto-acetate CoA transferase (thiophorase). “Ako ang nag saing, iba ang kumain.
Awww Emo much:(”

EICOSANOIDS  Autocrine (the same cell that secretes the hormone will be the one affected) and paracrine hormones (adjacent cells will be the ones that will be affected). Example: Prostaglandins, leukotrienes, thromboxanes, lipoxins Synthesized from o 20-carbon FA (AKA eicosanoic acids)  Arachindonic (4 double bonds)  series 2 prostaglandins (through cycleoxygenase pathway)  Timnodonate (5 double bonds)  Eicosapentaenoic acids  series 3 prostaglandins  Eicosatrienoate

 

After oxidation, cells form a lot of ketone bodies. And if cell is in starvation, the source of energy is from Acetyl CoA then the cell forms a lot of ketone bodies. Condensation of 2 moles of Acetyl CoA catalyzed by thiolase, with the release of coenzyme A forming acetoacetyl CoA. By means by synthetase and with another mole of Acetyl CoA, we will form 3-hydroxy-3-methylbutaryl CoA, with the release of acetyl CoA, we form the 1st ketone body—acetoacetate. Acetoacetate forms acetone.

Exert their effects through formation of cAMP.  They have to bind to the receptor in the plasma membrane which stimulates the G-protein which activates Adenylyl Cyclase, which forms cAMP out of ATP (cAMP protein dependent kinase cascade).

Physiological responses:        Negates inflammatory responses Produce pain and fever Regulate blood clotting Induce labor (for specific kinds) Have an effect on regulation of split weight cycle Inhibits gastric secretions Stimulate contraction of intestinal smooth muscles

PROSTAGLANDIN Identified in human semen and other cells Prostaglandin E and Prostaglandin F are initially discovered  Derived from hypothetical prostanoic acid  There are 3 major classes (depending on the structure): o Prostaglandin A – α,β- unsaturated ketones o Prostaglandin E- β- hydroxy ketones o Prostaglandin F- 1,3 dioles (2 hydroxy groups) Series 1 prostagalinds– there is 1 double bond outside the cyclicized region from eicosatrienoic acid or dihomogamma-linolenic acid Series 2 prostaglandins – there are 2 double bonds outside the cyclicized region from eicosatetraenoic acid or arachidonic acid  

Acetone   A volatile ketone body which is released through respiration. Its accumulation may also result to type 1 diabetes mellitus which has the propensity to develop ketoacidosis due to the absence of insulin synthesis.

3-hydroxy Butarate  Another ketone body

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Series 3 prostaglandins – there are 3 double bonds outside the cyclicized region from eicosapentaenoic acid or timnodonic acid Among the unsaturated fatty acids, there are 2 essential polyunsaturated acids: 1. linoleic acid (true essential) a. From this, we can synthesize dihomogamm-linolenic through a dehydrogenation reaction 2. linolenic acid

o

Cyclooxygenase activity is inhibited by aspirin (acetylsalicylic acid) and ibuprofen (Non-steroidal anti-inflammatory drugs)  2 isoforms of cyclooxygenase  COX1 – a constitutive enzyme (it’s always present whether or not there are substrates) o When blocked, GI bleeding and gastritis o Needed for the maintenance of integrity of the membranes of intestinal cells COX2 – an inducible enzyme o Made only in response through inflammatory initiators such as cytokines o The one that is blocked o Celecoxib are specific inhibitors Cyclooxygenase activity is inhibited by aspirin and ibuprofen (Nonsteroidal antiinflammatory drugs) 

LEUKOTRIENES o Catalysis through lipooxygenase o Series 3 from eicosatrienoate o Series 4 from arachidonic acid o Series 5 from eicosapentaenoate o

Pathway for Cyclooxygenase: Arachidonic acid is the most important precursor. o o Generation of Arachidonic acid through: o Reaction of phospholipase A2 which acts on carbon 2 of glycerol backbone. o Phospholipase A1 acts on carbon 1 of glycerol backbone o Phospholipase C acts on carbon 3 o Phospholipase D removes the base.  Ex: Enositol portion of phosphatidyl enositol is removed through this phospholipase D. o Diacylglycerol lipase o Diacylglycerol kinase Prostaglandin H2 synthase enzyme complex: o Possesses:  Cyclooxygenase activity  Aspirin inhibits this activity  Peroxidase activity

Structure of cyclooxygenase domain has 3 important amino acid residues 1. Tyrosine 3,8,5 a. Forms a radical in the process of forming prostaglandins b. If we give NSH, it will affect the tyrosine residue that prevents the formation of radical 2. Serine 5,30 a. Aspirin acetylates this, producing acetyl moiety which blocks the whole entry of arachidonic acid 3. Arginine 1,20 a. Forms an ion pair with substrate b. If arachidonic acid cannot enter, arginine cannot forms an ion pair with substrate Phospholipase A2 generates arachidonic acid o Hormones that stimulate angiotensin 2: o Bradykinin o Epinephrine o Thrombin Cortisol (principle corticosteroid) inhibits the activity of phospholipase A2

Phosphatidyl enositol through phospholipase A2  arachidonic acid + lysophospholipid Diacylglycerol kinase + phospholipase C  phosphatidic acid (+ phospholipase A2 arachidonic acid) Diacylglycerol lipase from 1,2 diacyl glycerol  arachidonic acid 2 major pathways of arachidonic acid metabolism: 1. Cyclooxygenase (Cyclic pathway) 2. Lipooxygenase (Linear pathway) Shows us how Arachidonic Acid  Prostaglandin o 2 activities:

o

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o o Prostaglandin dehydrogenase o Carbon attached to hydroxyl group is subsequently oxidized, and therefore inhibits prostaglandin o

WBC, Muscles, Lungs, Brain Actions:  Induces release of lysosomal enzymes  Promotes adhesion of WBC Message is transmited by the formation of 2nd messenger of cAMP

LIPOXINS   Family of congregated tetraenes Vasoactive and immunoregulatory effects

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Kung hindi clear yung image, please refer to the uploaded pdf file if it is available LIPOOXYGENASE PATHWAY o Peptidoleukotriene – mediates anaphylaxis o Leukotriene C4 o Leukotriene D4 o Leukotriene E4

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