Metabolism of Glycogen Peter A. Mayes, PhD, DSc, & David A. Bender, PhD BIOMEDICAL IMPORTANCE Glycogen is the major storage carbohydrate in animals, corresponding to starch in plants; it isa branched poly. mer of d-D-glucose. It occurs mainly in liver (up to 69%) and muscle, where it rarely exceeds 1%. However, be- cause of its greater mass, muscle contains about three to four times as much glycogen as docs liver (Table 18-1). ‘Muscle glycogen is a readily available source of glu- cose for glycolysis within the muscle itself. Liver glyco- gen functions to store and export glucose to maintain blood glucose between meals. After 12-18 hours of fasting, the liver glycogen is almost totally depleted. Glycogen storage diseases are a group of inherited dis- orders characterized by deficient mobilization of glyco- gen or deposition of abnormal forms of glycogen, lead- ing to muscular weakness or even death. GLYCOGENESIS OCCURS MAINLY IN MUSCLE & LIVER The Pathway of Glycogen Biosynthesis Involves a Special Nucleotide of Glucose (Figure 18-1) As in glycolysis, glucose is phosphorylated to glucose G-phosphate, catalyzed by hexokinase in muscle and glucokinase in liver. Glucose 6-phosphate is isomer- ized to glucose 1-phosphate by phosphoglucomutase. The enzyme itself is phosphorylated, and the phospho- group takes part in a reversible reaction in which glu- cose 1,6-bisphosphate is an intermediate. Next, glucose L-phosphate reacts with uridine triphosphate (UTP) to form the active nucleotide uridine diphosphate glu- cose (UDPGlc)* and pyrophosphate (Figure 18-2), catalyzed by UDPGle pyrophosphorylase. Pyrophos- = Other nucleoside diphosphate sugae compounds aze known, eg, ‘UDPGal. In addition, the same sugar may be linked co diferent nucleotides. For example, glacase may be linked eo uridine (as shown above) as well as to guanosine, thymidine, adenosine, of ¢: tidine nucleotides 145 phatase catalyzes hydrolysis of pyrophosphate to 2 mol of inorganic phosphate, shifting the equilibrium of the main reaction by removing one of its products Glycogen synthase catalyzes the formation of a gly- coside bond between C, of the activated glucose of UDPGle and C, of a terminal glucose residue of glyco: gen, liberating uridine diphosphate (UDP). A preexist- ing glycogen molecule, or “glycogen primer,” must be present t0 initiate this reaction. The glycogen primer may in turn be formed on a primer known as glyco- genin, which is a 37-kDa protein that is glycosylated on a specific tyrosine residue by UDPGlc. Further glu- cose residues are attached in the 14 position to make a short chain that is a substrate for glycogen synthase In skeletal muscle, glycogenin remains attached in the center of the glycogen molecule (Figure 13-15), whereas in liver the number of glycogen molecules is greater than the number of glycogenin molecules. Branching Involves Detachment of Existing Glycogen Chains ‘The addition of a glucose residue to a preexisting glyeo- gen chain, of “primer,” occurs at the nonreducing, outer end of the molecule so that the “branches” of the glycogen “tee” become elongated as successive 14 linkages are formed (Figure 18-3). When the chain has been lengthened to at least 11 glucose residues, branch- ing enzyme transfers a patt of the 194 chain (at least six glucose residues) to a neighboring chain co form a 1-36 linkage, establishing a branch point. The branches grow by further additions of 1>4-glucosyl units and further branching GLYCOGENOLYSIS IS NOT THE REVERSE OF GLYCOGENESIS BUT IS A SEPARATE PATHWAY (Figure 18-1) Glycogen phosphorylase catalyzes the rate-limiting seep in glycogenolysis by promoting the phosphorylytic cleavage by inorganic phosphate (phosphorylysis; cf hy-146 J CHAPTER 18 Glycogen (154 and 1-38 glucosyl units), (1-4 uc unt, pain 1 vse lo axcoam] . __O ® NcoSeN See acters Epinephrine auucan |* oe ayes Ed Uridine ENZYME i glucose (UDPGIc) Se) en, oP triphosphate (UTP) Glucose 1-phosphate foc pao pee f Figure 18-1. Pathway of glycogenesis and of glycogenolysis in the liver. Two high-energy phosphates are used in the incorporation of 1 mol of glucose into glycogen. ®, stimulation; ©, inhibition. Insulin decreases the level of cAMP only after it has been raised by glucagon or epinephrine—ie, it antagonizes their action. Glucagon is active in heart muscle but not in skeletal muscle. At asterisk: Glucan transferase and debranching enzyme ap- pear to be two separate activities of the same enzyme, Table 18-1. Storage of carbohydrate in postabsorptive normal adult humans (70 kg) drolysis) of the 1-94 linkages of glycogen to yield glu cose L-phosphate. The terminal glucosyl residues from the outermost chains of the glycogen molecule are re- Liver glycogen 4.0% Muscle ‘glycogen ore moved sequentially until approximately four glucose Pee tor Seose oie residues remain on cither side of a 1-96 branch (Figure 18-4). Another enzyme (0-[1-*4]a-1—+4] glucan transferase) transfers a trisaccharide unit from one ‘Ger weight 1800 branch to the other, exposing the 16 branch point "uscle mass35 ka Hydrolysis of the 196 linkages requires the de- "Total volume 10 branching enzyme. Further phosphorylase action canscHoH keuy ° J o- HOH ) 1H) } Ribose H H Hoon J Glucose Diphosphate Uridine Figure 18-2. Uridine diphosphate glucose (UDPGIc) then proceed. The combined action of phosphorylase and these other enzymes leads to the complete break- down of glycogen. The reaction catalyzed by phospho- glucomutase is reversible, so that glucose G-phosphate can be formed from glucose 1-phosphate. In liver (and kidney), but not in muscle, there is a specific enzyme, glucose-6-phosphatase, that hydrolyzes glucose G-phosphate, yielding glucose that is exported, leading to an increase in the blood glucose concentration, CYCLIC AMP INTEGRATES THE REGULATION OF GLYCOGENOLYSIS & GLYCOGENESIS ‘The principal enzymes conteolling glycogen metabo- lism—glycogen phosphorylase and glycogen synthase— are regulated by allosteric mechanisms and covalent modifications due to reversible phosphorylation and METABOLISM OF GLYCOGEN J 147 dephosphorylation of enzyme protein in response to hormone action (Chapter 9). Cyclic AMP (cAMP) (Figure 18-5) is formed from ATP by adenylyl cyclase at the inner surface of cell membranes and acts as an intracellular second messen- ger in response to hormones such as epinephrine, nor- epinephrine, and glucagon. cAMP is hydrolyzed by phosphodiesterase, so terminating hormone action. In liver, insulin increases the activity of phosphodiesterase, Phosphorylase Differs Between Liver & Muscle In liver, one of the serine hydroxyl groups of active phosphorylase a is phosphorylated. It is inactivated by hydrolytic removal of the phosphate by protein phos- phatase-I to form phosphorylase b. Reactivation re- quires rephosphorylation catalyzed by phosphorylase kinase. Muscle phosphorylase is distinct from that of liver. It is a dimer, each monomer containing I mol of pyridoxal phosphate (vitamin B,). It is present in two forms: phos- phorylase a, which is phosphorylated and active in either the presence or absence of AMP (its allosteric modi- fier); and phosphorylase b, which is dephosphorylaced and active only in the presence of AMP. This occurs during exercise when the level of 5’-AMP rises, providing, by this mechanism, fuel for the muscle. Phosphorylase ais the normal physiologically active form of the enzyme. cAMP Activates Muscle Phosphorylase Phosphorylase in muscle is activated in response to epi nephrine (Figure 18-6) acting via cAMP. Increasing, the concentration of cAMP activates cAMP-dependent © “C-labeled glucose residue 4 ut ® 3 gE EAE werctem SS 8 | $ = aa ie oa ? ? t Figure 18-3. The biosynthesis of glycogen. The mechanism of branching as revealed by adding “C-labeled glucose to the diet in the living animal and examining the liver glycogen at further intervals.148 J CHAPTER 18 PHOSPHORYLASE | [GLUCAN “TRANSFERASE DEBRANCHING ENZYME £-#)), Glucose residues joined by 0 J 1-94 glucosidie bonds Glucose residues joined by PO 146 alucosidi bonds Figure 18-4. Steps in glycogenolysis. protein kinase, which catalyzes the phosphorylation by ATP of inactive phosphorylase kinase b to active phosphorylase kinase a, which in curn, by means of a further phosphorylation, activates phosphorylase b to phosphorylase a Ca** Synchronizes the Activation of Phosphorylase With Muscle Contraction Glycogenolysis increases in muscle several hundred-fold immediately after the onset of contraction. This volves the rapid activation of phosphorylase by activa- sion of phosphorylase kinase by Ca’, the same signal as that which initiates contraction in response to nerve stimulation. Muscle phosphorylase kinase has four Nee 4 @ on 3/,5'-Adenylic acid (cyclic AMP; cAMP) Figure 18-5. types of subunits—o, B, y, and B—in a structure repre- sented as (O78), The 0: and f subunits contain serine residues that ate phosphorylated by cAMP-dependent protein kinase. The 8 subunit binds four Ca™* and is identical to the Ca**-binding protein calmodulin (Chapter 43). ‘The binding of Ca’* activates the cat- alytic site of the subunit while the molecule remains in the dephosphorylated b configuration, However, the phosphorylated a form is only fully activated in the presence of Ca". A second molecule of calmodulin, or ‘TpC (che structurally similar Ca’*-binding protein in muscle), can interact with phosphorylase Kinase, caus- ing further activation. Thus, activation of muscle con- action and glycogenolysis are carried out by the same Ca’*-binding protein, ensuring their synchronization, Glycogenolysis in Liver Can Be cAMP-Independent In addition to the action of glucagon in causing forma- tion of cAMP and activation of phosphorylase in liver, a1;-adrenergic receptors mediate stimulation of glyco- genolysis by epinephrine and norepinephrine. This in- volves a cAMP-independent mobilization of Ca” from mitochondria into the cytosol, followed by the stimulation of a Ca**/calmodulin-sensitive phosphory- lase kinase. cAMP-independent glycogenolysis is also caused by vasopressin, oxytocin, and angiotensin II act- ing through calcium or the phosphatidylinositol bis- phosphate pathway (Figure 43-7). Protein Phosphatase-1 Inactivates Phosphorylase Both phosphorylase a and phosphorylase kinase a are dephosphorylated and inactivated by protein phos- phatase-1. Protein phosphatase-1 is inhibited by a protein, inhibitor-1, which is active only after it has been phosphorylated by cAMP-dependent protein ki- nase, Thus, cAMP controls both the activation and in- activation of phosphorylase (Figure 18-6). Insulin re- inforces this effect by inhibiting the activation of phosphorylase b. It does this inditecdy by increasing uptake of glucose, leading to increased formation of glucose 6-phosphate, which is an inhibitor of phosphor- ylase kinase. Glycogen Synthase & Phosphorylase Activity Are Reciprocally Regulated (Figure 18-7) Like phosphorylase, glycogen synthase exists in cither a phosphorylated or nonphosphorylated state. However, unlike phosphorylase, the active form is dephosphory- lated (glycogen synthase a) and may be inactivated to6b Epinephrine B Receptor © Inactive Active adenylyl ‘adenylyl cyclase cyclase are camp seam Py = ae PROTEIN KINASE PROTEIN KINASE, PHOSPHORYLASE & ‘actve) ALVODULIN COMPONENT OF PHOSPHORYLASE| “KINASE 9 © PROSPHORYLASEKINASED] |. [PHosPHORMASEKINASE®]| <7 gop <\Y — nsutin PROTEN, ‘inset cal Taetve) PHOSPHATASE-t N_’ 6 PHOSPHORYLASE ADP lincivey PROTEN PHOSPHATASE-* 72 Figure 18-6. Control of phosphorylase in muscle. The sequence of reactions arranged as a cascade allows amplification of the hormonal signal at each step. (n = number of glucose residues; G6P, glucose 6-phosphate)150 / CHAPTER 18 Enineptrine B Receptor |) Inactive Aetve deny adenyhl cyclase ‘yeas [RiosmnoDieSTERRSE] ate cAMP S-AMP PHOSRIORIASE @ “ena a aa aie cau SEE cap SEE Moenr Grew Nase MOieWeNase are aycoser in Innibtort (inactive) TADIQDUNCDEPENDENT SOE Russe are CLraGEKETITINSE] Gf [evcoELSMMTASE @ inactive) Catt factve) Tasuln > GP soe | POTN, POSTS oy yeogenn H.0' i bee rot prsshate 7 gtoER, fate © Figure 18-7. Control of glycogen synthase in muscle (n = number of glucose residues). The sequence of reac- tions arranged in a cascade causes amplification at each step, allowing only nanomole quantities of hormone to ‘cause major changes in glycogen concentration. (GSK, glycogen synthase kinase-3, 4, and -5; G6P, glucose 6-phosphate.) glycogen synthase b by phosphorylation on serine residues by no fewer than six different protein kinases Two of the protein kinases are Ca’*/calmodulin- dependent (one of these is phosphorylase kinase). An- other kinase is cAMP-dependent protein kinase, which allows cAMP-mediated hormonal action to inhibic glycogen synthesis synchronously with the activation of glycogenolysis, Insulin also promotes glycogenesis in muscle ar the same time as inhibising glycogenolysis by raising glucose G-phosphate concentrations, which stimulates the dephosphorylation and activation of glycogen synthase. Dephosphorylation of glycogen syn thase b is carried out by protein phosphatase-1, which is under the control of cAMP-dependent protein ki- REGULATION OF GLYCOGEN METABOLISM IS EFFECTED BY ABALANCE IN ACTIVITIES BETWEEN GLYCOGEN SYNTHASE & PHOSPHORYLASE (Figure 18-8) Not only is phosphorylase activated by a rise in concen: tration of cAMP (via phosphorylase kinase), but glyco- gen synthase is at the same time converted to the inactive form; both effects are mediated via cAMP- dependent protein kinase. ‘Thus, inhibition of gly- cogenolysis enhances net glycogenesis, and inhibition of elycogencsis enhances net glycogenolysis. Furthermore,METABOLISM OF GLYCOGEN / 151 Epinephrine PIOSPHODIESTERASE fiver, muscle) o : biter inititor4 Glucagon an? SrAMP Inhibitors phosphate tier ® ‘sivcoaen PHOSPHORYLASE SYNTHASE RNASE pRoTEN eaoreN PHOSPHATASE: PHOSPHATASE: \ , \ © NC ‘stvcoaen PHOSPHORYLASE SYNTHASE 8 KINASE & ao Glycogen vorcie — Syeegen PHOSPHORYLASE PHOSPHORYLASE \ Glucose 1-phosphate 4IN Glucose (ver) Glucose Lactate (muscle) Figure 18-8. PROTEIN PHOSPHATASE.1 le Coordinated control of glycogenolysis and glycogenesis by cAMP-dependent protein ki- nase, The reactions that lead to glycogenolysis as a result of an increase in cAMP concentrations are shown with bold arrows, and those that are inhibited by activation of protein phosphatase-1 are shown as broken arrows. The reverse occurs when cAMP concentrations decrease as a result of phosphodiesterase activity, leading to glycogenesis. the dephosphorylation of phosphorylase a, phosphory- Jase kinase a, and glycogen synthase b is catalyzed by a single enzyme of wide specificity—protein phos- phatase-1. In turn, protein phosphatase-1 is inhibited by cAMP-dependent protein kinase via inhibitor-1 Thus, glycogenolysis can be terminated and glycogenesis can be stimulated synchronously, or vice versa, because both processes are keyed to the activity of cAMP-depen: dent protein kinase. Both phosphorylase kinase and glycogen synthase may be reversibly phosphorylated in more than one site by separate kinases and phosphatases. These secondary phosphorylations modify the sensiciviey of the primary sites to phosphorylation and dephos- phorylation (multisite phosphorylation). What is more, they allow insulin, via glucose 6-phosphate eleva- tion, to have effects that act reciprocally to those of cAMP (Figures 18-6 and 18-7) CLINICAL ASPECTS Glycogen Storage Diseases Are Inherited “Glycogen storage disease” is a generic term to describe a group of inherited disorders characterized by deposi- tion of an abnormal type or quantity of glycogen in the tissues. The principal glycogenoses are summarized in Table 18-2. Deficiencies of adenylyl kinase and cAMP-dependent protein kinase have also been re152 J CHAPTER 18 Table 18-2. Glycogen storage diseases. _Slycogenosis a: Typel Von Gierke’s disease i Type Pompe's disease Type Limit dextrinosis, Forbes’ or Con's disease i Type lv ‘Amylopectinosis, Andersen's disease TypeV Myophosphorylase deficiency, McaArdle's syndrome : TypeV Hers disease Type Vi Tarui’s disease Type Vil kinase Cause of Disorder Deficiency of glucose- | Deficiency of lysosomal a-1-4-and | 16-glucosidase (acid maltase) | Absence of debranching enzyme bsence of branching enzyme ‘Absence of muscle phosphorylase Deficiency of liver phosphorylase | Deficiency of phosphofructokinase “in muscle and erythrocytes ! Deficiency of liver phosphorylase Characteristics Liver cells and renal tubule cells loaded | with glycogen. Hypoglycemia lactic- acidemia, ketosis, hyperlipemia, pphosphatas Fatal, accumulation of glycogen in lyso- | somes, heart failure | Accumulation of a characteristic | branched polysaccharide, | Accumulation of a polysaccharide hav- | ing few branch points. Death due to | cardiac oliver failure i Diminished exercise tolerance; muscles have abnormally high glycogen con- tent (2.5-4.196) Little or no lactate in blood after exercise. High glycogen content in liver, ten- 'y toward hypoglycemia ‘sor type V but aso possibility of he- molytic anemia Asfor type Vl ported. Some of the conditions described have bene- fited from liver transplantation, SUMMARY + Glycogen represents the principal storage form of carbohydrate in the mammalian body, mainly in the liver and muscle. + In the liver, its major function is to provide glucose for extrahepatic tissues. In muscle, it serves mainly as a ready source of metabolic fuel for use in muscle Slycogen is synthesized from glucose by the pathway of glycogenesis. Iris broken down by a separate path- way known as glycogenolysis. Glycogenolysis leads to glucose formation in liver and lactate formation in muscle owing to the respective presence or absence of glucose-6-phosphatase. lic AMP integrates the regulation of glycogenoly- sis and glycogenesis by promoting the simultancous activation of phosphorylase and inhibition of glyco- gen synthase. Insulin acts reciprocally by inhibiting elycogenolysis and stimulating glycogenesis. + Inherited deficiencies in specific enzymes of glycogen metabolism in both liver and muscle are the causes of glycogen storage diseases. REFERENCES Bollen M, Keppens S, Stalmans W: Specific features of glycogen. rictabolism in the liver. Biochem | 1998:336:19. ‘Cohen P: The tole of protein phosphorylation in the hormonal control of enzyme activity. Fur J Biochem 1985;151:439. Ercan N, Gannon MC, Nutall FQ: Incorporation of glycogenin into a hepatic proteoglycogen alter oral glucose administra sion. J Biol Chem 1994:269:22328, Geddes R: Glycogen: a metabolic viewpoint. Bioscience Rep 1986:6:415, McGanty JD ct al: From dietary glucose co liver glycogen the full circle round. Annu Rey Nutt 1987,7:51 Meléadee-Hevia E, Waddell TG, Shelton ED: Optimization of rmoleculst design in the evolution of metabolism: the glyco: gen molecule. Biochem J 1993:295:477, [Raz I, Kate A, Spencer MK: Epinephrine inhibits insulin-mediated lycogenesis but enhances elycolysit in human skeletal mus dle, Am J Physiol 1991;260:8430. Seriver CR etal (editors): The Metabolic and Molecular Bases of In- berited Direase, 8th ed. MeGraw-Hill, 2001 Shulman GI, Landau BR: Pathways of glycogen repletion, Physiol Rev 1992,72:1019. Villar-Palasi C: On the mechanism of inactivation of muscle glyco- gen phosphorylate by insulin. Biochim Biophys Acca 1994; 1224:384,