UNIT II:

Intermediary Metabolism

Glycogen metabolism
Figure 11.1. Glycogen synthesis and degradation shown as a part of the essential
reactions of energy metabolism (see Figure 8.2, p. 90, for a more detailed view of
the overall reactions of metabolism)..
 Overview
• A constant source of blood glucose is an absolute requirement for
human life
• Glucose is the greatly preferred energy source for the brain, & the
required energy source for cells with few or no mitoch., e.g.,
mature RBCs
• Glucose is also essential as an energy source for exercising
muscle, where it is the substrate for anaerobic glycolysis.
• Blood glucose can be obtained from 3 primary sources: diet,
degradation of glycogen, & gluconeogenesis.
• Dietary intake of gluc & gluc precursors, e.g., starch,
monosacch’s, & disacch’s, is sporadic and, depending on diet, is
not always a reliable source of gluc.
• In contrast, gluconeogenesis can provide sustained synthesis of
gluc, but it is somewhat slow in responding to falling blood gluc
level
• Therefore, body has developed mechanisms of storing a supply of
glucose in a rapidly mobilizable form i.e., glycogen.
• In absence of a dietary source of gluc, this cpd is rapidly
released from liver & kidney glycogen. Similarly, muscle
glycogen is extensively degraded in exercising muscle to
provide that tissue with an important energy source
• When glycogen stores are depleted, specific tissues
synthesize gluc de novo, using aa’s from body’s proteins
as primary source of carbons for gluconeogenic
pathway.
II. Structure and function of glycogen
- The main stores of glycogen in the body are found in
skeletal muscle & liver, although most other cells store
small amounts of glycogen for their own use
- Function of muscle glycogen is to serve as a fuel
reserve for synthesis of ATP during muscle contraction
- That of liver glycogen is to maintain blood glucose
conc., particularly during early stages of fast
A. Amounts of liver and muscle glycogen
- ~ 400 g of glycogen make up one to two percent of the
fresh weight of resting muscle, & ~ 100 g of glycogen
make up to 10% of the fresh weight of a well-fed adult
liver. What limits production of glycogen at these levels
is not clear.
- However, in some glycogen storage diseases, the
amount of glycogen in liver and/or muscle can be
significantly higher
Figure 10.2. Functions of muscle
and liver glycogen.
B. Structure of glycogen
- Glycogen is a branched-chain homo-polysaccharide
made exclusively from α-glucose
- The primary glycosidic bond is an α(1→4) linkage.
- After an av. of 8-10 glucosyl residues, there is a branch
containing an α(1→6) linkage.
- A single molecule of glycogen can have a molecular
mass of up to 108 daltons.
- These molecules exist in discrete cytoplasmic granules
that contain most of the enz’s necessary for glycogen
synthesis & degradation
Figure 11.3. Branched structure of glycogen, showing a-1,4 and a-1,6
linkages.
C. Fluctuation of glycogen stores
- Liver glycogen stores increase during the well-fed state
& are depleted during a fast
- Muscle glycogen is not affected by short periods of
fasting ( a few days) & is only moderately decreased in
prolonged fasting (weeks).
- Muscle glycogen is synthesized to replenish muscle
stores after they have been depleted, e.g., following
strenuous exercise.
Note: synthesis & degradation of glycogen are processes
that go on continuously. Differences b/w rates of these 2
processes determine levels of stored glycogen during
specific physiologic states
III. Synthesis of glycogen (glycogenesis)
- Glycogen is synthesized from molecules of α-D-glucose. The process
occurs in cytosol, and requires energy supplied by ATP (for
phosphorylation of gluc) & uridine triphosphate (UTP)
A. Synthesis of UDP-glucose
- α-D-gluc attached to UDP is the source of all of glucosyl residues
that are added to the growing glycogen molecule
- UDP-gluc is synthesized from glucose-1-P & UTP by UDP-glucose
pyrophosphorylase
- The high-energy bond in pyrophosphate (PPi), the 2nd product of the
reaction, is hydrolyzed to 2 inorganic phosphates (Pi) by
pyrophosphatase, which ensures that synthesis of UDP-gluc
proceeds in direction of UDP-gluc production
Note: G-6-P is converted to G-1-P by phosphoglucomutase. G-1,6-BP
is an obligatory intermediate in this reaction
Figure 11.4. The structure of UDP-glucose.
B. Synthesis of a primer to initiate glycogen synthesis
- Glycogen synthase is responsible for making α (1→4) linkages in
glycogen. This enz can’t initiate chain synthesis using free gluc as
an acceptor of a molecule of gluc from UDP-gluc. Instead, it can
only elongate already existing chains of gluc
- Therefore, a fragment of glycogen can serve as a primer in cells
whose glycogen stores are not totally depleted
- In the absence of a glycogen fragment, a protein, called glycogenin,
can serve as an acceptor of gluc residues
- Side chain hydroxyl group of a specific Tyr serves as the site at
which the initial glucosyl unit is attached
- Transfer of first few molecules of gluc from UDP-gluc to glycogenin
is catalyzed by glycogenin itself, which can then transfer additional
glucosyl units to the growing α (1→4)-linked glucosyl chain
- This short chain serves as an acceptor of future gluc residues
Note: glycogenin stays associated with & is found in center of
completed glycogen molecule
Figure 11.5. Glycogen synthesis.
C. Elongation of glycogen chain by glycogen synthase
- Elongation of glycogen chain involves transfer of gluc
from UDP-gluc to the non-reducing end of growing chain,
forming a new glycosidic bond b/w the anomeric
hydroxyl of C-1 of activated gluc & C-4 of accepting
glucosyl residue
Note: “non-reducing end” of a CHO chain is one in which
anomeric C of terminal sugar is linked by a glycosidic
bond to another cpd, making terminal sugar “non-
reducing”.
- The enz responsible for making α (1→4) linkages in
glycogen is glycogen synthase
Note: UDP released when the new α (1→4) glycosidic bond
is made can be converted back to UTP by nucleoside
diphosphate kinase (UDP + ATP ↔ UTP + ADP)
Figure 11.6. Interconversion of glucose 6-phosphate and glucose 1-
phosphate by phosphoglucomutase.
D. Formation of branches in glycogen
- If no other synthetic enz’s acted on the chain, resulting
structure would be a linear molecule of glucosyl residues
attached by α (1→4) linkages.
- Such a cpd is found in plant tissues, & is called amylose.
In contrast, glycogen has branches located, on av., 8
glucosyl residues apart, resulting in a highly branched,
tree-like structure that is far more soluble than
unbranched amylose
- Branching also increases the # of non-reducing ends to
which new glucosyl residues can be added (and also,
from which these residues can be removed), thereby
greatly accelerating the rate at which glycogen synthesis
& degradation can occur, & dramatically increasing the
size of the molecule
1. Synthesis of branches:
- Branches are made by action of “branching enzyme”,
amylo-α (1→4) → α (1→6)-transglucosidase. This enz
transfers a chain of 5 to 8 glucosyl residues from non-
reducing end of glycogen chain [breaking α (1→4) bond]
to another residue on the chain and attaches it by an α
(1→6) linkage
- Resulting new, non-reducing end, as well as the old non-
reducing end from which the 5 to 8 residues were
removed, can now be elongated by glycogen synthase

2. Synthesis of additional branches:

- After elongation of these two ends has been accomplished
by glycogen synthase, their terminal 5 to 8 glucosyl
residues can be removed & used to make further
branches
IV. Degradation of glycogen (glycogenolysis)

• The degradative pathway that mobilizes stored glycogen

in liver & skeletal muscle is not a reversal of the
synthetic reactions. Instead a separate set of cytosolic
enz’s is required.
• When glycogen is degraded, the primary product is G-1-
P, obtained by breaking α (1→4) glycosidic bonds. In
addition, free gluc is released from each α (1→6)-linked
glucosyl residue
A. Shortening of chains
- Glycogen phosphorylase sequentially cleaves the α
(1→4) glycosidic bonds b/w the glucosyl residues at the
non-reducing ends of glycogen chains by simple
phosphorolysis until 4 glucosyl units remain on each
chain before a branch point
Note: this enz contains a molecule of covalently bound
pyridoxal phosphate that is required as a coenzyme
- Resulting structure is called a limit dextrin, &
phosphorylase can’t degrade it any further
Cleavage of an α (1→ 4)-glycosidic bond.
B. Removal of branches
- Branches are removed by 2 enzymatic activities. 1st
oligo-α(1→4)→α (1→4)-glucan transferase removes the
outer 3 of the 4 glucosyl residues attached at a branch. It
next transfers them to the non-reducing end of another
chain, lengthening it accordingly. Thus, an α(1→4) bond
is broken and an α(1→4) bond is made.
- Next, the remaining single gluc residue attached in an
α(1→6) linkage is removed hydrollytically by amylo-
α(1→6)-glucosidase activity, releasing free gluc.
Note: both the transferase & glucosidase are domains of a
single polyp molecule, the ‘debranching enzyme”.
- The glucosyl chain is now available for degradation by
glycogen phosphorylase until 4 glucosyl units from next
branch are reached
Figure 11.8
Glycogen degradation,
showing some of the
glycogen storage
diseases. (Continued on
next page.)
Figure 11.8 (Continued )
C. Conversion of G-1-P to G-6-P
- G-1-P, produced by glycogen phosphorylase, is
converted in the cytosol to G-6-P by
phosphoglucomutase, a reaction that produces G-1,6-BP
as a temporary but essential intermediate
- In liver, G-6-P is translocated into ER by glucose 6-
phosphate translocase. There it is converted to glucose
by glucose 6-phosphatase, the same enz used in last
step of gluconeogenesis
- Resulting glu is then transported out of ER to cytosol.
Hepatocytes release glycogen-derived gluc into blood to
help maintain blood gluc levels until gluconogenic
pathway is actively producing gluc
Note: in muscle, G-6-P can’t be dephosphorylated because
of a lack of glucose-6-phosphatase. Instead, it enters
glycolysis, providing energy needed for muscle
contraction
D. Lysosomal degradation of glycogen
- A small amount of glycogen is continuously degraded by
lysosomal enz, α(1→4)-glucosidase (acid maltase).
Purpose of this pathway is unknown
- However, a deficiency of this enz causes accumulation
of glycogen in vacuoles in the cytosol, resulting in the
serious glycogen storage disease type II (Pompe
disease)
V. Regulation of glycogen synthesis & degradation

• Because of importance of maintaining blood gluc levels,

synthesis & degradation of its glycogen storage form are
tightly regulated
• In liver, glycogen synthesis accelerates during periods
when the body has been well fed, whereas degradation
accelerates during periods of fasting.
• In skeletal muscle, glycogen degradation occurs during
active exercise, & synthesis begins as soon as the
muscle is again at rest
• Regulation of glycogen synthesis & degradation is
accomplished on two levels
• First, glycogen synthase & glycogen phosphorylase are
allosterically controlled.
• Second, the pathways of glycogen synthesis &
degradation are hormonally regulated
Note: regulation of glycogen synthesis & degradation is
extremely complex, involving many enz’s (e.g., protein
kinases & phosphatases), calcium, & enz inhibitors,
among others
A. Allosteric regulation of glycogen synthesis & degradation
- Glycogen synthase & glycogen phosphorylase respond
to levels of metabolites & energy needs of cell. It is
logical, therefore, that glycogen synthesis is stimulated
when substrate availability & energy levels are high,
whereas glycogen degradation is increased when energy
levels & available gluc supplies are low
1. Regulation of glycogen synthesis & degradation in the
well fed state:
- In the well fed state, glycogen synthase is allosterically
activated by G-6-P when it is present in elevated conc’s.
in contrast, glycogen phosphorylase is allosterically
inhibited by G-6-P, as well as ATP, a high-energy signal
in cell.
Note: in liver, gluc also serves as an allosteric inhibitor of
glycogen phosphorylase
Figure 11.9. Allosteric regulation of glycogen synthesis and degradation
in A. Liver, and B. Muscle.
2. Activation of glycogen degradation in muscle by calcium
- During muscle contraction, there is a rapid & urgent need for ATP,
the energy for which is supplied by muscle’s stores of glycogen.
- Nerve impulses cause memb depolarization, which in turn promotes
Ca2+ release from sarcoplasmic reticulum into sarcoplasm of
muscle cells
- Ca2+ binds to calmodulin, one of a family of small, calcium-binding
proteins
Note: calmodulin is the most widely distributed of these proteins, & is
present in virtually all cells
- Binding of 4 molecules of Ca2+ to calmodulin triggers conformational
change such that activated Ca2+-calmodulin complex binds to &
activates protein molecules, often enz’s, that are inactive in absence
of this complex
- Thus, calmodulin functions as an essential subunit of many complex
proteins. One such protein is phosphorylase kinase, which is
activated by Ca2+-calmodulin complex without need for the kinase
to be phosphorylated by cAMP-dependent protein kinase
- When muscle relaxes, Ca2+ returns to sarcoplasmic reticulum &
phosphorylase kinase becomes inactive
Note: phosphorylase kinase is maximally active in exercising muscle
when it is both phosphorylated & bound to Ca2+
Figure 11.10. Calmodulin mediates many
effects of intracellular calcium.
3. Activation of glycogen degradation in muscle by AMP
- Muscle glycogen phosphorylase is active in presence of
high AMP conc’s that occur in muscle under extreme
conditions of anoxia & ATP depletion
- AMP binds to the inactive form of glycogen
phosphorylase, causing its activation without
phosphorylation
B. Activation of glycogen degradation by cAMP-directed
pathway
- Binding of hormones, e.g., glucagon & epinephrine, to
memb receptors signals the need for glycogen to be
degraded, either to elevate blood gluc levels or to
provide energy for exercising muscle
1. Activation of protein kinase:
- Binding of glucagon or epinephrine to their specific CM
receptors  cAMP-mediated activation of cAMP-
dependent protein kinase.
- This enz is a tetramer, having 2 regulatory (R) & 2
catalytic (C) subunits.
- cAMP binds R subunit dimer, releasing individual C
subunits that are active
Note: when cAMP removed, inactive tetramer R2C2, is
again formed
2. Activation of phosphorylase kinase:
- Phosphorylase kinase exists in 2 forms: an inactive “b”
form & an active “a” form
- Active cAMP-dependent protein kinase phosphorylates
inactive form of phosphorylase kinase  activation
Note: phosphorylated enz can be inactivated by hydrolytic
removal of its P by protein phosphatase 1. This enz is
activated by a kinase-mediated signal cascade initiated
by insulin

3. Activation of glycogen phosphorylase:

- Glycogen phosphorylase also exists in 2 forms: the
dephosphorylated, inactive “b” form & phosphorylated,
active “a” form.
- Active phosphorylase kinase phosphorylates glycogen
phosphorylase a, which then begins glycogen
breakdown
- Phosphorylase a is converted to phosphorylase b by
hydrolysis of its P by protein phosphatase 1.
Note:
- when gluc is bound to glycogen phosphorylase a, thus
signaling that glycogen degradation is no longer
required, the complex becomes a better substrate for
protein phosphatase 1.
- In addition, when muscle glycogen phosphorylase b is
bound to glucose, it can’t be allosterically activated by
AMP
- In the muscle, insulin indirectly inhibits the enz by
increasing uptake of gluc, leading to an increased level
of G-6-P, a potent allosteric inhibitor of glycogen
phosphorylase
Figure 11.11. Stimulation and inhibition of glycogen degradation.
4. Summary of regulation of glycogen degradation:
- Cascade of reactions listed above result in glycogen
degradation
- The large # of sequential steps serves to amplify the
effect of the hormonal signal, i.e., a few hormone
molecules binding to their receptors result in a # of
protein kinase molecules being activated that can each
activate many phosphorylase kinase molecules
- This causes production of many active glycogen
phosphorylase a molecules that can degrade glycogen
C. Inhibition of glycogen synthesis by a cAMP-directed
pathway
- The regulated enz in glycogen synthesis is glycogen
synthase.
- It also exists in 2 forms, the “a” form which is not
phosphorylated & is the most active form, & “b” form,
which is phosphorylated & inactive.
- Glycogen synthase a is converted to “b” form by
phosphorylation at several sites on the enz, with the
level of inactivation is proportional to its degree of
phosphorylation
- This conversion process is catalyzed by several different
protein kinases that are regulated by cAMP or other
signaling mechanisms
Note: protein kinase C, a Ca2+ & phospholipid-dependent
protein kinase, also phosphorylates glycogen synthase.
Neither protein kinase A nor C directly phosphorylates
glycogen phosphorylase
- Binding of glucagon or epinephrine to hepatocyte
receptors, or of epinephrine to muscle cell receptors,
results in the activation of adenylyl cyclase, mediated by
G-protein
- This enz catalyzes synthesis of cAMP, which activates
cAMP-dependent protein kinase A
- Protein kinase A then phosphorylates & thereby
inactivates glycogen synthase
- Glycogen synthase “b” can be transformed back to
synthase “a” by protein phosphatase 1, which removes P
groups hydrolytically
Figure 11.12. Hormonal regulation of glycogen synthesis. [Note: In contrast to
glycogen phosphorylase, glycogen synthase is inactive if phosphorylated.]
VI. Glycogen storage diseases
- These are a group of genetic diseases that result from a
defect in an enz required for glycogen synthesis or
degradation
- They result either in formation of glycogen that has an
abnormal structure, or in the accumulation of excessive
amounts of normal glycogen in specific tissues as a
result of impaired degradation
- A particular enz may be defective in a single tissue, such
as liver, or the defect may be more generalized, affecting
liver, muscle, kidney, intestine, & myocardium
- Severity of glycogen storage diseases (GSDs) ranges
from fatal in infancy to mild disorders that are not life-
threatening
 Summary
• Main stores of glycogen in body are found in skeletal
muscle, where they serve as a fuel reserve for synthesis
of ATP during muscle contraction, & in liver, where
glycogen is used to maintain blood glucose conc,
particularly during early stages of a fast
• Glycogen is a highly branched polymer of α-D-glucose.
The primary glycosidic bond is an α (1→4) linkage. After
~ 8-10 gluc residues, there is a branch containing an α
(1→6) linkage.
• UDP-gluc, building block of glycogen, is synthesized
from G-1-P & UTP by UDP-glucose pyrophosphorylase
• Gluc from UDP-glucose is transferred to the non-
reducing ends of glycogen chains by glycogen synthase,
which makes α (1→4) linkages
• Branches are formed by amylo-α(1→4) → α(1→6)-
transglucosidase, which transfers a chain of 5-8 glucosyl
residues from the non-reducing end of glycogen chain
(breaking an α(1→4) linkage), and attaches it with an
α(1→6) linkage to another residue in the chain
• Glycogen phosphorylase cleaves the α(1→4) bonds b/w
glucosyl residues at the non-reducing ends of glycogen
chains, producing G-1-P. it requires pyridoxyl phosphate
as a coenz.
• This sequential degradation continues until 4 glucosyl
units remain on each chain before a branch point. The
resulting structure is called a limit dextrin.
• Oligo-α(1→4)→α(1→4)-glucan transferase [common
name, glucosyl (4:4) transferase] removes the outer 3 of
the 4 glucosyl residues attached at a branch, & transfers
them to the non-reducing end of another chain where
they can be converted to G-1-P by glycogen
phosphorylase.
• Next, the remaining single gluc residue attached in an
α(1→6) linkage is removed hydrolytically by the amylo-
α(1→6)-glucosidase activity, releasing free gluc.
• G-1-P is converted to G-6-P by phosphoglucomutase. In
the muscle, G-6-P enters glycolysis. In liver, the P is
removed by glucose-6-phosphatase, releasing free gluc
that can be used to maintain blood gluc levels at
beginning of a fast
• A deficiency of the phosphatase causes glycogen
storage disease type 1 (Von Gierke disease). This
disease results in an inability of liver to provide free gluc
to body during a fast. It affects both glycogen
degradation & last step in gluconeogenesis
• Glycogen synthase & glycogen phosphorylase are
allosterically regulated. In well-fed state, glycogen
synthase is activated by G-6-P, as well as ATP
• In liver, gluc also serves as an allosteric inhibitor of
glycogen phosphorylase
• Ca2+ is released from sarcoplasmic reticulum during
exercise. It activates phosphorylase kinase in the muscle
by binding to enz’s calmodulin subunit. This allows the
enz to activate glycogen phosphorylase, thereby causing
glycogen degradation
• Glycogen synthesis & degradation are reciprocally
regulated by the same hormonal signals, namely, an
elevated insulin level results in overall increased
glycogen synthesis & decreased degradation, whereas
elevated glucagon (or epinephrine) level causes
increased glycogen degradation & decreased synthesis.
• Key enz’s are phosphorylated by a family of protein
kinases, some of which are cAMP-dependent (a cpd
increased by glucagon and epinephrine). Phosphate
groups are removed by protein phosphatase 1 (activated
when insulin levels are elevated).
Figure 11.13. Key concept map for glycogen metabolism in liver.