Lecture 30: Fatty Acid Catabolism I I. Four major roles for fatty acids a. Fuel i. Storage as triacylglycerols b.

Component of membrane bilayers c. Building blocks for phospholipids d. Precursors for hormones and signaling molecules Chemistry and modular nature of machinery in fatty acid metabolism a. Analogous with non-ribosomal peptide synthesis Advantages/disadvantages of fats a. Adv: high energy content (highly reduced) b. Dis: insoluble in water Human heart and liver a. Fatty acid oxidation = 80% of energetic needs i. Under all physiological conditions Electrons removed from fatty acids pass through a. Acetyl CoA -> TCA -> oxidative phosphorylation i. Generating ATP b. In liver converted to ketone bodies i. Serves as fuel for brain, etc. when glucose low Tri acyl glycerols = neutral fats a. Insoluble i. Must be emulsified before water soluble enzymes in intestine can digest Cells can obtain fatty acid fuels from 3 sources a. Fats consumed in diet b. Fats stored in cells (lipid droplets) c. Fats synthesized in one organ and exported to another Vertebrates a. Obtain fat in diet b. Mobilize fats stored in adipocytes Triacylglycerols a. Major source of energy for heart and liver Dietary fats solubilized by bile salts and absorbed through intenstinal wall a. Ingested triacylglycerols converted from insoluble macroscopic fat particles i. Into soluble fine microscopic micelles b. Bile salts (biological detergents) i. Glycocholate ii.Taurocholic acid c. Micelle structure i. Cone shaped d.

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Lipase action catalyzes breakdown of triacylglycerols

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a. b. These triacylglycerol derivatives cross intenstinal mucosa i. Re-converted to triacylglycerols 1. Packed into lipoproteins called chylomicrons Chylomicrons a. Aggregates of fatty acids triacylglycerols + apo-lipoproteins i. apo- implies separate (lipid-free form) ii.Together form lipoprotein particles 1. Hydrophobic lipids at core 2. Hydrophilic protein side chains and lipid head groups at surface b. Apolipoprotein part of lipoproteins contains recognition domains i. Control transport of the particles ii.APO C-II (apolipoprotein C-II) 1. Lipid uptake from the intestinal mucose -> lymphatic system -> bloodstream c. Then carried to muscle and adipose tissue i. Metabolized by lipoprotein lipase (extracellular enzyme) 1. Activated by APO-CII 2. Transported into tissue where fatty acids are B-oxidation (oxidation of fatty acids) a. Occurs in 3 stages

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c. Stage 1 i. Oxidative removal of successive two-carbon units 1. Forms 1 acetyl-CoA for each two-carbon unit a. Ex: 16-carbon palmitate => 8 acetyl-coA 2. Each removal requires removal of four hydrogen atoms by dehydrogenases 3. Overall reaction

4. d. Stage 2 i. Acetyl-CoA oxidized -> CO2 1. Via citric acid cycle (mitochondrial matrix) e. Stage 3 i. First two stages of fatty acid oxidation => reduced NADH and FADH2 ii.In stage 3, donate electrons to mitochondrial electron-transfer chain 1. Leads to ADP-> ATP a. Thus energy released by fatty acid oxidation => ATP B-oxidation of poly-unsaturated fatty acids (multiple double bonds) a. If two cis-bonds i. First cis bond changed by isomerase 1. But after b-oxidation, another cis-bond remains

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b. c. After Acyl-CoA dehydrogenase acts on this intermediate i. Forming double bond between C2-C3 ii.This new product (dienoyl CoA) not substrate for the hydratase d.

e. Solution: reductase used to reduce unwanted double bond (bottom to top)

i. ii.Isomerization then follows as before (enoyl CoA isomerase) Lecture 31

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Overall ATP production of glycolysis + TCA cycle a. 1 NADH = 2.5 ATP (Ox Phos) b. 1 FADH2 = 1.5 ATP (Ox Phos) c. Glucose (C6) => 32 ATP i. 5.3 ATP / C d. Palmitate (C16) => 108 ATP i. 6.8 ATP / C Peroxisomes also carry out B-oxidation a. Same pathway as mitochondria except for 1st Omega (w) oxidation by cytochrome P450 a. Oxidation of carbon most distant from carboxyl group b. Useful when B-oxidation defective

c. i. Results in double-ended fatty acids that can go through Boxidation on both ends 1. Yielding dicarboxylic acids (C-6 to C-10) d. Omega oxidation in liver and kidney i. Results in significant levels of C-6 to C-10 dicarboxylates in urine Ketone bodies a. Uses Acetyl-CoA when OAA is limiting (Fasting, diabetes, etc.) i. Acetyl-CoA + OAA = substrates for TCA cycle ii.Allows continued oxidation of fatty acids in liver when acetylCoA cant be metabolized by TCA cycle Metabolic disease states related to fatty acid oxidation a. MCAD (medium chain acyl-CoA dehydrogenase deficiency) i. Low levels of urinary ketone bodies ii.High levels of dicarboxylic acids from omega oxidation 1. Acyl-CoA dehydrogenase deficiency = B-oxidation defective

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a. Low levels of acetyl-CoA => low levels of ketone bodies b. Diabetic ketosis/acidosis or starvation i. High levels of urinary ketone bodies ii.Starvation 1. Gluconeogenesis depletes TCA intermediates a. Acetyl-CoA converted to ketone bodies iii.Diabetes 1. Low insulin levels = tissue cant uptake glucose 2. Under these conditions a. Malonyl-CoA levels fall (starting material for fatty acid synthesis) i. Inhibition of carnitine acyltransferase relieved b. Fatty acids enter mitochondria => acetyl-CoA 3. Acetyl-CoA cant enter TCA b/c of gluconeogenesis a. Extra ketone bodies formed = lower blood pH => acidosis b. Too much ketone bodies in blood/urine => ketosis Lecture 32: Amino Acid Catabolism, Urea Cycle I. Metabolic conditions when amino acids are being oxidized a. Normal AA turnover b. After rich protein meal i. And when little protein synthesis c. Starvation or diabetes when carbohydrates not being metabolized Focus on fate of nitrogen a. Amino acid groups have amino group that must be metabolized i. Unlike carbohydrates, fatty acids b. This amino group (nitrogen) metabolism links urea cycle and TCA cycle i. In human protein metabolism ii.And maintenance of nitrogen homeostasis In hepatocyte cytosol (liver cells) a. Amino groups transferred to alpha-ketoglutarate => glutamate i. Glutamate moved to mitochondria 1. Converted to ammonia In most tissues a. Excess ammonia converted to glutamine and imported into liver mitochondria In muscle a. Excess amino groups transferred to alanine i. Transported to liver

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b. Dietary protein is enzymatically degraded to amino acids a. Starts in stomach where pepsin secreted (acidic conditions) i. Cuts at certain AAs, shortens peptides b. Near pancreatic duct pH raises to 7 c. Goes to intestines i. Peptide degradation finishes

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d. Urea cycle linked to TCA cycle a. Urea located in cytosol TCA cycle located in mitochondrial matrix b. Cycles linked by cytosolic isozymes of enzymes of TCA cycle i. Ex: fumarase and malate dehydrogenase 1. Present as isozymes in cytosol

a. Fumarate generated can be converted to malate in cytosol i. Can be further metabolized in cytosol ii.Or brought to mitochondira for TCA cycle ii.The products of these isozymes can easily be used in both cycles

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c. Activity of urea cycle regulated at two levels a. Long term i. Rate of urea cycle enzyme synthesis 1. Higher in high-protein diet and starvation (eating muscle protein) a. Urea cycle = disposing nitrogen (amino group) b. Short term i. Allosteric regulation 1. First enzyme in pathway = carbomyl phosphate synthetase I a. Allosterically activated by N-acetylglutamate i. Which synthesized from acetyl-CoA and glutamate by n-acetylglutamate synthase 1. Purely regulatory enzyme ii.Levels of N-acetylglutamate dependent on levels of acetyl-CoA ad glutamate iii.Also dependent on arginine (activator of nacetylglutamate synthase) 1. Arginine = activator of urea cycle Metabolic disease linked to urea metabolism a. Patients cant tolerate protein rich diets b. Carbonyl phosphate synthetase deficiency i. Treated with benzoate or phenyacetate 1. Gives path to dump ammonia

ii. 1. First forms intermediate with CoA (coenzyme A) a. Requires energy in form of ATP-> AMP b. S-CoA replaced with amino group of amino acid i. The amino-acid complex is excreted c. Arginosuccinase deficiency

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i. Carbons of amino acids enter TCA at different points a. As different substrates

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b. Ketogenic amino acids a. Seven amino acids that degraded entirely or in part to i. acetoacetyl-CoA and/or acetyl-CoA 1. then converted to acetoacetate then -> acetone and Bhydroxybutyrate glucogenic amino acids a. amino acids that degraded to pyruvate i. alpha-ketoglutarate, succinyl-CoA, fumarate, and/or oxaloacetate 1. can be converted to glucose and glycogen (gluconeogenesis) b. Some amino acids both glucogenic and ketogenic Cofactors in one-carbon transfer important in AA catabolism a. Biotin i. Transfers one- carbon in most oxidized state: CO2 b. Tetrahydrofolate i. Transfers one-carbon group in intermediate oxidation states: HCOH 1. Sometimes methyl ii.Can accept different oxidation states of one carbon unit 1. CH3, CH2OH, CHO c. S-adenosylmethionine i. Transfers most reduced state: CH3

ii. iii.N5-methyltetrahydrofolate transfers CH3 group to homocysteine 1. Forming methionine a. Joins adenosine => S-adenosyl-methionine i. Transfers CH3 to another molecule