BIOCHEMISTRY

GLYCOGENOLYSIS
Dra. Catherine L. Co-Reportoso | October 31, 2018 LE3 TRANS1

OUTLINE  GLYCOGEN BREAKDOWN OVERVIEW:

I. Review of Glycogen III. Glycogen Storage Disease
 Glycosidic bond between two glucosyl units is cleaved
Metabolism IV. Review Questions
in glycogen and converted to Glucose-1-Phosphate
II. Glycogenolysis V. References
A. Liver Glycogen  Enzyme: Glycogen Phosphorylase
B. Muscle Glycogen  Reaction: Phosphorolysis (hydrolyzing chemical bonds
C. Steps in Clycogenesis while adding a phosphate)
D. Regulation  Glucose-1-phosphate  Glucose-6-phosphate
LEARNING OBJECTIVES  Enzyme: Phosphoglucomutase
At the end of the lecture, the student should be able to:  Location: Cytosol
1. Discuss glycogenolysis  G-6-P Free Glucose
2. Explain the regulation of glycogenolysis
 Enzyme: Glucose-6-phosphatase
3. Discuss the different glycogen storage diseases
 Location: ER lumen
I. REVIEW OF GLYCOGEN METABOLISM
 Reaction: Hydrolysis
 Glycogenesis
 the formation of glycogen from glucose A. LIVER GLYCOGEN
 occurs in liver and muscle cells when glucose and ATP are  100g of glycogen = 10% of well-fed adult liver [2021C Trans]
present in relatively high amounts  After 12 to 18 hrs of fasting, liver glycogen is almost totally
 one ATP and one UTP are required for every glucose unit depleted [Harper’s 31st ed.]
incorporated into the polymeric branched structure of  Main source of fuel for body tissues 8-12 hours after meal
glycogen during the synthesis of glycogen
 Liver stored glycogen are mobilized for distribution to other
 Glycogenolysis tissues [2021A Trans]
 Takes place in the cells (cytosol) of muscle and liver tissues  Responds to high or low levels of blood glucose
in response to hormonal and neural signals
 GLUT2 transports glucose to the bloodstream
 Plays an important role in fight-or-flight response and
 Regulation
regulation of blood glucose levels
 Insulin (increase glycogen synthesis) and Glucagon
(increase glycogenolysis) hormones counteraction
Structure and Function of Glycogen
 Epinephrine as stimulator of glycogenolysis
B. MUSCLE GLYCOGEN
 The role of muscle glycogen is to provide a source of
glucose-6-phosphate for glycolysis in response to the need
for ATP for muscle contraction [Harper’s 31st ed.]
 Lack of glucose-6-phosphatase = cannot create free glucose
(for energy)
 Glycogen is abundant in Type II Muscle fibers (White muscle
fibers) – greater capacity for glycogenolysis and glycolysis but
for shorter periods of time.

Figure 1: Structure of Glycogen [Tutorvista.com] Table 1. Glycogen source abundance in Type I and Type II Muscle Fibers

 Major carbohydrate storage in humans and animals Type I Muscle fiber Type II Muscle fiber
 Made up of a chain of α-D glucose units linked (Red Fibers) (White Fibers)
 linearly by α (1, 4) glycosidic bonds Mitochondria
 branches connected by α (1, 6) glycosidic bonds Abundant Low
Myoglobin
 Terminal residues are the sites of action of glycogen Oxidative capacity High Low
phosphorylase and synthase
Glycogen and
 Muscle glycogen – for future energy uses; storage reserve for Glycolytic activity
Low High
ATP synthesis Postural and Axial
 Muscle glycogen provides readily available source of Muscle Appendicular
muscles
glucose-1-phosphate for glycolysis within the muscle itself Main source of GLYCOGEN
TAGs
 Liver glycogen – serve to replenish blood glucose levels during energy Creatine PO4
fasting states
*Remember Muscle Physiology:
II. GLYCOGENOLYSIS Type 1 - One Slow Red Ox Type 2 – Two Fast White
Sugar(Glycogen)
 Glycogenolysis in NOT the reverse of Glycogenesis, but a
separate pathway, separate set of cytosolic enzyme [Harper’s 31st
 Regulation
ed.]
 Insulin inhibits glycogen breakdown
 Due to the presence of Glycogen phosphorylase  Muscle cells lack receptors for glucagon [Mowshowitz, 2011][2021A
 rate-limiting step in glycogenolysis Trans]
 phosphorolytic cleavage of the 1 → 4 linkages of glycogen  Stimulators of Glycogenolysis: Epinephrine (from adrenal
to yield Glucose-1-phosphate glands), Glucagon (from pancreas), presence of AMP
 Different isoenzymes in liver, muscle and brain

L.E. # 3 - Trans # 1 Group G: Robeniol, Robles, A. Rodriguez, S. Rodriguez, Romero 1 of 6

 C. STEPS IN GLYCOGENOLYSIS  cleaves α -1,6 glycosidic bond of the last glucose residue,
releasing a free glucose molecule (in G1P form)
Phosphorolysis and shortening of chains  Branched glycogen becomes more linear and is now
available again for degradation by glycogen
phosphorylase until four glucosyl units from the next
branch are reached [Lipincott Illustrated Biochemistry]
 Action: Hydrolysis
 Both enzymes have separate activities of a single protein in two
catalytic sites
 Both enzymes convert the branched glycogen into a more linear
form to be readily cleaved by glycogen phosporylase
Figure 2. Action of glycogen phosphorylase on the glycogen residues linked by
α-1, 4-glycosidic linkage Isomerization of G-1-P to G-6-P
 Enzyme: glycogen phosphorylase  Site: Cytosol
 Key enzyme in cleaving α 1,4- glycosidic bond on the
glycogen residue by adding an orthophosphate to yield G-
1-P
 Coenzyme: pyridoxal phosphate (PLP)
 Covalently bound to phosphorylase
 Used to form carbocation (carbonium ion)
 Action: shortening of glycosyl chain from the non-reducing end
by Phosphorolysis
 Cleaves α 1,4- glycosidic bond between the glucosyl
residues at non-reducing ends of glycogen chain.
 Yields G-1-P molecules which are readily converted to G-6-
P via Phosphoglucomutase
 Cleavage stops only when 4 glucose units are left before
the α 1,6- branch point
 Does not cleave α 1,6- glycosidic bond
 Limit dextrin: resulting structure when phosphorylase stops
cleaving
 Dextrin cannot be degraded any further by glycogen
phosphorylase
 Rate limiting step
 No water is used
Figure 4. Isomerization of G-1-P to G-6-P
Debranching step/ Glycogen Remodeling
→ Branches of dextrin removed by 2 enzymic activities of a single  Enzyme: Phosphoglucomutase (PGM)
bifunctional protein  Exchanges phosphoryl group with substrate
→ Removal of glucose residues at the branch point  Reversible
 Action: PGM exchanges a phosphoryl group with the
substrate G-1-P (reverse of the 1st step in glycogenesis)
 Phosphoryl group is transferred from serine residue (of PGM)
to the C6 hydroxyl group of glucose
 Temporary (but essential) Intermediate:
G-1,6-bisphosphate
 Dephosphorylated PGM accepts phosphoryl group from C1
of G-1,6-bisphosphate
 Product: G-6-P

Fates of G-6-P
1. May enter the ff. pathways [2022B Biochem trans]
Pathway Produce
Glycolysis ATP needed for muscle contraction
Pentose Phosphate Ribose
Figure 3. Debranching Step
Pathway (PPP) NADPH
→ Enzyme: Debranching enzymes Gluconeogenesis Free glucose
 Oligo- α (1 → 4) →α (1 → 4)-glucan transferase or
4-α-D-glucanotransferase
2. In the liver, G-6-P is further converted to glucose via
 Transfers outer three of the four glucosyl residues in the
glucose-6-phosphatase
limit branch to the main branch
3. In skeletal muscle, enzyme glucose-6-phosphatase is
 an α (1→4) bond is broken and an α(1→4) bond is made
absent; it enters glycolysis. Hence, muscular glycogen
and the enzyme functions as a 4:4 transferase [Lipincott
cannot contribute to blood glucose replenishing.
Illustrated Biochemistry]
 exposes single/last glucose residue attached to the main
Dra. Reportoso: Skeletal muscle functions to produce energy
branch via α -1,6 glycosidic bond and α 1,6 glycosidic
(straight to glycolysis) while Liver functions to produce glucose for
bond is also exposed
blood sugar level (G6P converted to glucose)
 amylo- α (1 → 6)-glucosidase

Biochemistry Glycogenolysis 2 of 6
Lysosomal Degradation of Glycogen  Regulation of synthesis and degradation is accomplished on 2
levels
 Hormonally Regulated (covalent)
 Enzymes are hormonally regulated to meet the needs of
the body as a whole
 Allosterically Regulated
 Enzymes are allosterically regulated to meet the needs of
a particular tissue
 Inhibition of glycogenolysis enhances net glycogenesis;
Inhibition of glycogenesis enhances net glycogenolysis
 In fasting state, increase ↑ glucagon and decrease ↓ insulin
(+) initiate a cAMP-directed phosphorylation cascade
 Phosphorylation of glycogen phosphorylase activates it
(active [a] form)
 Phosphorylation of glycogen synthase inactivates it
(inactive [b] form)
As a consequence, active glycogen phosphorylase and inactive
glycogen synthase promotes glycogenolysis (glycogen
degradation) and inhibits glycogenesis (glycogen synthesis)

Hormonal Regulation
Figure 5. Lysosomal Degradation of Glycogen Activation of Glycogenolysis
 Enzyme: acid maltase or α (1 → 4) glucosidase
 1-3% of glycogen is continuously degraded by acid maltase
 Pompe disease (Type II GSD): accumulation of glycogen in the
body due to the absence of acid maltase

Hydrolysis of G-6-P to glucose

Figure 6. Glucose Transport in liver

 Enzyme: Glucose-6-phosphatase (absent in muscles)

 Location: ER Lumen (Kidney and Liver) Figure 7. Activation of Glycogenolysis
 G-6-P Translocase (T1): transports G-6-P to ER lumen  Mechanism: Activation of Glycogenolysis by cAMP directed
 Glucose Transporter (T2): transports glucose back to pathway
cytosol  Regulatory Enzyme: Glycogen Phosphorylase
 Pi Transporter (T3): transports Pi to cytosol  Mechanism:
 GLUT2: transports glucose from cytosol to capillaries 1. Glucagon or epinephrine binds to receptors (specific
(blood stream) hepatocyte GPCRs or myocyte GPCR for epinephrine)
2. Binding results in the activation of adenylyl cyclase
D. REGULATION OF GLYCOGENOLYSIS 3. Adenylyl cyclase catalyzes the synthesis of cAMP which
 Because of the importance of maintaining blood glucose levels, activates cAMP-dependent protein kinase A (PKA)
the synthesis and degradation of its glycogen stores are tightly 4. PKA phosphorylates inactive phosphorylase kinase B
regulated to active phosphorylase kinase A
5. Active phosphorylate kinase A phosphorylates (activates)
Pathway Liver Muscle
Glycogen Phosphorylase (inactive (b) glycogen
Glycogenesis Well-fed state At rest phosphorylase is activated by Phosphorylase Kinase A to
Glycogenolysis Fasting state Active (exercise) active (a) glycogen phosphorylase)
6. Activation of Glycogen Phosphorylase stimulates
glycogenolysis
 2 principal regulatory enzymes
 Glycogen Phosphorylase is dephosphorylated (inactivated)
 Glycogen Phosphorylase
by protein phosphatase-1 [Harper’s, 2012]
 Rate limiting enzyme
 Exists in two forms:
Inhibition of Glycogenesis
 active [a] form (phosphorylated) = R (relaxed) State
(Refer to the appendix for Hormonal Regulation of Glycogen Synthesis)
 inactive [b] form (dephosphorylated) = T (tensed) State
 Mechanism: Inhibition of Glycogenesis by cAMP directed
 Glycogen Synthase
pathway
 Exists in two forms:
 Regulatory Enzyme: Glycogen Synthase
 active [a] form (dephosphorylated)
 Mechanism:
 inactive [b] form (phosphorylated)
Biochemistry Glycogenolysis 3 of 6
 1. Six different protein kinases act on Glycogen Synthase
[Harper’s, 2012]
2. Phosphorylation of Glycogen Synthase by the different
protein kinases inactivates it (active (a) glycogen
synthase is inactivated to inactive (b) glycogen synthase)
3. Inactivation of Glycogen Synthase inhibits glycogenesis
Allosteric Regulation
 Aside from hormonal signals, glycogen synthase and glycogen
phosphorylase respond to the levels of metabolites and energy
needs of the cell
 Glycogenesis is stimulated when glucose and energy levels are
high whereas glycogenolysis is increased when glucose and
energy levels are low
Figure 7. Allosteric control of glycogen metabolism (Lippincott)
 In a fed state, glycogen phosphorylase is in the inactive
dephosphorylated form (glycogen phosphorylase b) due to the
effect of insulin. This action prevents the body from producing
more glucose, hence, preventing hyperglycemia.
 In a fed state, glucose would enter the allosteric sites,
changing the conformation of the enzyme (phosphatase) and
exposing the phosphate groups
 Insulin activates protein phosphatase-1, which
dephosphorylates glycogen phosphorylase into its inactive
form glycogen phosphorylase b → (-) glycogenolysis
 Effect of Glucose-6-Phosphate
 In a well-fed state or under resting conditions, there is an
accumulation of G-6-P both in the liver and in the muscles
 G-6-P allosterically activates (dephosphorylates) glycogen
synthase → (+) glycogenesis
 G-6-P allosterically inactivates (dephosphorylates) glycogen
phosphorylase → (-) glycogenolysis
 Effect of Calcium
 During muscular contraction, there is an increased demand
for ATP → Ca2+ is released at the ER and Ca2+ binds with
calmodulin
 Calcium-calmodulin complex convert phosphorylase
kinase b to phosphorylase kinase a → (+) glycogenolysis
 Glycogen conversion to G6P → G6P enters glycolysis →
ATP generation
 Effect of AMP
 High AMP concentrations activate muscle glycogen
phosphorylase (glycogen phosphorylase a) under
conditions of ATP depletion → (+) glycogenolysis
→glycogen conversion to G6P → G6P enters glycolysis
→ATP generation
 AMP binds to glycogen phosphorylase b causing its
activation without having to be phosphorylated
III. GLYCOGEN STORAGE DISEASES
 Generic term to describe inherited disorders that are
characterized by an abnormality in the quantity of glycogen
in tissues, including its failure to mobilize glycogen.
 These diseases are primarily due to a defect on an enzyme that
is required in the glycogen synthesis/degradation.
Biochemistry Glycogenolysis
 Impairment of Glycogen Metabolism disallows the body to make
enough glucose and distribute it, or not be able to use glucose
as a form of energy.
 Formation of abnormal glycogen or accumulation can result to
impairment of tissue’s function or structure (Hepatomegaly /
Muscle Weakness).
 Only Type II GSD (Pompe’s Disease) is the only one that
includes lysosome impairment; characterized by accumulation
of abnormal amounts of carbohydrates or lipids due to their
decreased lysosome degradation.
GLYCOGEN STORAGE DISEASES (GSD)
Type 0
 Biochemical Basis: Glycogen Synthase Deficiency
 Clinical Manifestations: Low lactate and alanine, postprandial
hyperglycemia and high lactic acid in blood, fasting resulting in
hypoglycemia, ketotic hypoglycemia when ceasing night-time
feeds, seizures can occur, muscle cramps, glycosuria
 Diagnostic Modalities: Genetic testing
Type Ia (Von Gierke’s Disease)
 Biochemical Basis: Glucose-6-Phosphatase Deficiency
 Clinical Manifestations: Low lactate and alanine, postprandial
hyperglycemia and high lactic acid in blood, fasting resulting in
hypoglycemia, ketotic hypoglycemia when ceasing night-time
feeds, seizures can occur, muscle cramps, glycosuria
 Diagnostic Modalities: Enzyme Assay on Liver Biopsy
Type II (Pompe’s Disease)
 Biochemical Basis: Lysosomal Maltase (Alpha 1,4
glucosidase) Deficiency
 Clinical Manifestations: Enlarged heart, myopathy, muscular
dystrophy-like, engorged lysosomes.
 Diagnostic Modalities: test for creatine kinase, GAA assay,
muscle biopsy
Type III (Cori’s Disease)
 Biochemical Basis: liver and muscle Debranching Enzyme
Deficiency
 Clinical Manifestations: muscular weakness, hypoglycemia,
enlarged liver, ketoacidosis, no rise in blood glucose when
glucagon was administered
 Diagnostic Modalities: Liver Biopsy
Type V (McArdle’s Disease)
 Biochemical Basis: Muscle Phosphorylase Deficiency
 Clinical Manifestations: cramps, lifelong history of fatigability,
high levels of creatine kinase, lactate dehydrogenase, and
myoglobulin
 Diagnostic Modalities: Muscle biopsy
Dra. Reportoso’s Mnemonics:
1. ABCDEFG
Andersen’s (Type IV) ⇒ Branching enzyme
Cori’s (Type IIIa) ⇒ Debranching enzyme
Fanconi - Bickel (Type XI) ⇒ GLUT2
2. Mcardle’s (Type V) ⇒ Muscle phosphorylase
3. Hers (Type VI) ⇒ Hepatic phosphorylase
(Refer to the appendix for GSD Table)
IV. REVIEW QUESTIONS
1. Due to sleep deprivation during exam week, CR decided to
go home, sleep and woke up at 6am. He decided to walk to
UERM (30mins walk) and climbed the stairs. What is the
predominant glycosyl residue in his muscle that will serve as
a fuel store?
a. Free glucose
b. Glucose-1-phosphate
4 of 6
 c. Glucose-6-Phosphate
d. UDP glucose 2

For nos 2-5 refer to the following choices:

a. Type Ia
b. Type II
c. Type IIIa
d. Type IV
e. Type V
f. Type VI

2. Accumulation of glycogen in lysosomes of liver, heart and

muscle; juvenile form of myopathy
3. Liver glucose-6-PO4 deficiency
4. Exercise intolerance
5. Also known as Andersen disease
6. ↑ Limit Dextrin
7. Liver phosphorylase Deficiency
Answers: C, B, A, E, D, C, F

V. REFERENCES
 Lipincott’s Illustrated Review for Biochemistry
 Harper’s Illustrated Biochemistry
 Batch 2021 Transcriptions
 Dra. Reportoso’s lecture
 Lecture notes
 Structure of Glycogen https://chemistry.tutorvista.com/organic-
chemistry/glycogen.html
 2022B Biochem trans
 http://oregonstate.edu/instruct/bb450/fall14/stryer7/21/figure_21_19.jpg

Biochemistry Glycogenolysis 5 of 6
 APPENDIX

Type Name Deficient enzyme Clinical Features

Type XII Aldolase A Exercise intolerance and weakness
following febrile illnesses*
Type XIII β-enolase Rapidly progressive exercise
intolerance and myalgia, chronically
elevated serum CK*
Type XIV Congenital Phosphoglucomutase Hepatopathy, bifid uvula, malignant
disorder of hyperthermia, hypogonadotropic
glycosylation hypogonadism, growth retardation,
type It hypoglycemia, myopathy, dilated
cardiomyopathy, and cardiac arrest
Type XV Glycogenin Muscle weakness associated with
the depletion of glycogen in skeletal
muscle and cardiac arrhythmias
associated with the accumulation of
abnormal storage material in the
heart*
*clinical features were based on one reported case as these GSD types are rare

Biochemistry Glycogenolysis 6 of 6