FUNCTIONS OF GLYCOGEN Glycogen synthesized and stored in cytosolic granules in the liver and muscle.

cle. Liver glycogen maintain blood glucose concentration within normal range during early fasting. Muscle glycogen fuel reserve for the synthesis of ATP during muscle contraction.

GLYCOGENOLYSIS 4 ENZYMATIC ACTIVITIES FOR EFFICIENT BREAKDOWN OF GLYCOGEN One to degrade glycogen Two to remodel glycogen so it remains a substrate for degradation One to convert the product of glycogen breakdown into a form suitable for further metabolism. Cleaves substrate by phosphorolysis addition of Pi Product is Glucose 1-PO4 Sequential removal of glycosyl residues from non-reducing ends (the one with free 4-OH groups) Pi splits the glycosidic link between C-1 of terminal residue and C-4 of the next residue. Pi specifically: 1. Cleaves the bond between C-1 carbon atoms and glycosidic oxygen atom. 2. Retains the alpha configuration.

GLYCOGENOLYSIS PHOSPHOROLYTIC CLEAVAGE OF TERMINAL 1,4 GLYCOSIDIC BONDS AT NON-REDUCING END REMOVAL OF BRANCHES BY A DEBRANCHING ENZYME has 2 enzyme activities a.) Glucose 4:4 transferase ([1-4] -> [1-4] glucan transferase) b.) Amylo 1,6 glucosidase

 SYNTHESIS OF UDP-GLUCOSE FORMATION OF GLYCOGEN PRIMER ELONGATION OF GLYCOGEN CHAINS (AMYLOSE CHAIN FORMATION) FORMATION OF BRANCHES

AMINO ACID CARBON SKELETONS

Amino acids, when deaminated, yield -keto acids that, directly or via additional reactions, feed into major metabolic pathways (ex, Krebs Cycle).

Amino acids are groups into 2 classes, based on whether or not carbon skeletons can be converted to glucose:

Glucogenic Ketogenic

Carbon skeletons of glucogenic amino acids are degraded to: Pyruvate or A 4-C or 5-C intermediate of Krebs Cycle. These are precursors for gluconeogenesis. Glucogenic amino acids are the major carbon source for gluconeogenesis when glucose levels are low. They can also be catabolized for energy, or converted to glycogen or fatty acids for energy storage. Carbon skeletons of ketogenic amino acids are degraded to: Acetyl-CoA, or Acetoacetate Acetyl CoA, and its precursor acetoacetate, cannot yield net production of oxaloacetate, the gluconeogenesis precursor. For every 2-C acetyl residue entering Krebs Cycle, 2C leave as CO2. Carbon skeletons of ketogenic amino acids can be catabolized for energy in Krebs Cycle, or converted to ketone bodies or fatty acids. They cannot be converted to glucose.

The 3-C -keto acid pyruvate is produced from alanine, cysteine, glycine, serine and threonine. Alanine deamination via Transaminase directly yields pyruvate.

Serine is deaminated to pyruvate via Serine Dehydratase. Glycine, which is also product of threonine catabolism, is converted to serine by a reaction involving tetrahydrofolate.

The 4-C Krebs Cycle intermediate oxaloacetate is produced from aspartate and aspargine. Aspartate transamination yields oxaloacetate. Aspartate is also converted to fumarate in Urea Cycle. Fumarate is converted to oxaloacetate in Krebs Cycle. Aspargine loses the amino group from its Rgroup by hydrolysis catalyzed by Asparaginase. This yields aspartate, which can be converted to oxaloacetate. Ex., by transamination. The 4-C Krebs intermediate succinyl-CoA is produced from isoleucine, valine and methionine. Propionyl-CoA, an intermediate on these pathways, is also a product of -oxidation of fatty acids with an odd number of C atoms.

Proprionyl-CoA is carboxylated to methymalonyl-CoA. A racemase yields the L-isomer essential to the subsequent reaction. Methymalonyl-CoA Mutase catalyzes a molecular rearrangement: the branched C chain of methymalonyl-CoA is converted to the linear C chain of succinylCoA. The carboxyl that is in ester linkage to the thiol of coenzyme A is shifted to an adjacent carbon atom, with opposite shift of a hydrogen atom.

Coenzyme B12, a derivative of vitamin B12 (cobalamin), is the prosthetic group of Methymalonyl-CoA Mutase. Methyl group transfers are also carried out by B12 (cobalamin) in mammalian cells. Methy-B12, (methycobalamin), with a methyl axial ligand substituting for the deoxyadenosyl moiety of coenzyme B12, is an intermediate of such transfers. BRANCHED CHAIN AMINO ACIDS Branched chain amino acids initially share in part a common pathway. Branched Chain -Keto Acid Dehydrogenase (BCKDH) is a multi-subunit complex homologous to Pyruvate Dehydrogenase complex. Genetic deficiency of BCKDH is called Maple Syrup Urine Disease (MSUD).

High concentrations of branched chain keto acids urine give it a characteristic odor.

The 5-C Krebs Cycle intermediate ketoglutarate is produced from arginine, glutamate, glutamine, histidine and proline. Glutamate deamination via Transaminase directly yielding ketoglutarate.

Glutamate deamination by Glutamate Dehydrogenase also directly yields -ketoglutarate. THF exists in various forms, with single-C units of varying oxidation state, bonded at N5 or N10, or bridging between them. In these diagrams N10 with R is -aminobenzoic acid, linked to a chain of glutamate residues. The cellular pool of THF includes various forms, produced and utilized in different reactions.

N3-formimino-THF is involved in the pathway for degradation of histidine. Reactions using THF as donor of a single-C unit include synthesis of thymidylate, methionine, f-methionine-tRNA, etc.

In the pathway of histidine degradation, N-formiminoglutamate is converted to glutamate by transfer of the formimino group to THF, yielding N5-formimino-THF.

Histidine is first converted to glutamate. The last step in this pathway involves the cofactor tetrahydrofolate. Tetrahydrofolate (THF), which has a pteridine ring, is a reduced form of the B vitamin folate. Within a cell, THF has an attached chain of several glutamate residues, linked to one another by isopeptide bonds involving the R-group carboxyl. AROMATIC AMINO ACIDS Aromatic amino acids phenylalanine and tyrosine are catabolized to fumarate and acetoacetate.

Hydroxylation of phenylalanine to form tyrosine involves the reductant tetrahydrobiopterin. Biopterine, like folate, has a pteridine ring. Dihydrobiopterin is reduced to tetrahydrobiopterin by electron transfer from NADH. Thus NADH is secondarily the e- donor for conversion of phenylalanine to tyrosine. Tyrosine is a precursor for synthesis of melanins and of epinephrine and norephinephrine. High [phenylalanine] inhibits Tyrosine Hydroxylase, on the pathway for synthesis of the pigment melanin from tyrosine. Individuals with phenylketonuria have light skin and hair color.

Pheylalanine Hydroxylase includes a non-heme iron atom and its active site. X-ray crystallography has shown the following are ligands to the iron atom: His N, Glu O & water O, (fe shown in spacefill & ligands in ball & stick). O2, tetrahydropbiopterin, and the iron atom in the ferrous (Fe++) oxidation state participate in the hydroxylation. O2 is thought to react initially with the tetrahydropbiopterin to form a peroxy intermediate. Genetic deficiency of Phenylalaline Hydroxylase leads to the disease phenylketonuria. Phenylalanine & phenylpyruvate (the product of phenylalanine deamination via transaminase) accumulate in blood and urine. Mental retardation results unless treatment begins immediately after birth. Treatment consists of limiting phenylalanine intake to levels barely adequate to support growth. Tyrosine, an essential nutrient for individuals with phenylketonuria, must be supplied in the diet.

Or methionine may be regenerated from homocysteine by methy transfer from N5-methyl-tetrahydrofolate, via an enzyme that uses B12 as prosthetic group. Another pathway converts homocysteine to glutathione.

In various reactions, S-adenosylmethionine (SAM) is a donor of diverse chemical groups including methylene, amino, ribosyl and aminoalkyl groups, and a source of 53-deoxyadenosyl radicals. But SAM is best known as methyl group donor.

Examples: S-adenosylmethionine as methyl group donor: Methylation of bases in tRNA Methylation of cystosine residues in DNA Methylation of norepinephrine - > epinephrine

Conversion of glycerophospholipid Phosphatidyl ethanolamine -> phosphatidylcholine via methyl transfer from SAM. Enzymes involved in formation and utilization of Sadenosylmethionine are particularly active in liver. Liver has important roles in synthetic pathways involving methylation reactions and in regulation of blood methionine.

METHYL GROUP DONORS Methyl group donors in synthetic reactions include:

Methyl-B12 S-adenosylmethionine (SAM) N5-methyl-tetrahydrofolate (N5-methyl-THF)