GLYCOGENESIS

DR.SUDHANSHU SHEKHAR
ASSOCIATE PROFESSOR
DEPT . OF BIOCHEMISTRY
 GLYCOGEN- INTRODUCTION

• Glycogen is a readily mobilized storage form

of glucose.
• It is a very large, branched polymer of glucose
residues that can be broken down to yield
glucose molecules when energy is needed.
• Most of the glucose residues in glycogen are
linked by α-1,4-glycosidic bonds.
• Branches at about every tenth residue are
created by α-1,6-glycosidic bonds
 GLYCOGEN CHEMISTRY
• Glycogen is present in the cytosol in the form of
granules ranging in diameter from 10 to 40 nm.
• It has a molecular mass of 107 Da and consists of
polysaccharide chains, each containing about 13
glucose residues.
• The chains are either branched or unbranched and
are arranged in 12 concentric layers.
• The branched chains (each has two branches) are
found in the inner layers and the unbranched
chains in the outer layer.
• Muscle- synthesis of ATP,
• Liver- Maintenance of blood glucose.
 GLUCOSE
POLYSACCHARIDE

GLYCOGENIN

The highly branched structure of glycogen provides a large number

of sites for glycogenolysis, permitting rapid release of glucose 1-
phosphate for muscle activity.
• It is stored mainly in liver muscle
. and Brain.
• The liver content of glycogen is greater than that of
muscle,
• Since the muscle mass of the body is considerably
greater than that of the liver, about three-quarters of
total body glycogen is in muscle
• STORAGE OF CARBOHYDRATE IN 70 KG PERSON
PERCENTAGE TISSUE BODY
OF TISSUE WEIGHT CONTENT
WEIGHT
LIVER GLYCOGEN 5.0 1.8 kg 90 g

MUSCLE 0.7 35 kg 245 g

GLYCOGEN
EXTRACELLULAR 0.1 10 L 10 g
GLUCOSE
REASONS FOR STORING GLYCOGEN AS A FUEL

• Glycogen serves as a buffer to maintain blood-

glucose levels.
• Glucose is virtually the only fuel used by the
brain, except during prolonged starvation.
• The glucose from glycogen is readily mobilized
and is therefore a good source of energy for
sudden, strenuous activity.
• Unlike fatty acids, the released glucose can
provide energy in the absence of oxygen and can
thus supply energy for anaerobic activity
 GLYCOGENESIS
• Glycogenesis is the synthesis of glycogen from
glucose.
• Glycogenesis mainly occurs in muscle and
liver.
• Muscle glycogen provides a readily available
source of glucose for glycolysis within the
muscle itself.
• Liver glycogen functions to store and export
glucose to maintain blood glucose between
meals.
Liver And Muscle Glycogen
Glycogen synthesis requires the following:
1. UDP glucose : used as source of glucose
2. Glycogenin protein:
-37 Kda protein
- it is glycosylated at specific tyrosine residue by UDP-
Glucose
- further, glucose residue are added to it by α 1-4 linkage
to form a short chain which acts as a glycogen primer.
3. Glycogen Synthase- it adds glucose units to a pre
existing glycogen primer ( Glycogenin ) by α 1-4 linkage
4. Branching enzyme : it creates a branch point by adding
glucose by α 1-6 linkage
 STEPS OF GLYCOGENESIS

A. Synthesis of UDP-glucose
B. Synthesis of a primer to initiate glycogen
synthesis
C. Elongation of chain by glycogen synthase
D. Formation of branches
8

8
 A. Synthesis of UDP-glucose

• Glucose is phosphorylated to Glu-6-PO4 by

Hexokinase in muscle and Glucokinase in liver.
• Glu-6-PO4 by mutase is converted into Glu-1-PO4
• Glucose 1-phosphate reacts with uridine
triphosphate (UTP) to form the active nucleotide
uridine diphosphate glucose (UDPGlc) and
pyrophosphate. The reaction is catalyzed by
UDPGlc pyro phosphorylase.
• Spontaneous hydrolysis of the ~P bond in PPi (P~P)
drives the overall reaction
 .
• Cleavage of PPi is the only energy cost for glycogen
synthesis (one ~P bond per glucose residue).
• UDP-glucose, the glucose donor in the biosynthesis
of glycogen, is an activated form of glucose

UDP-glucose
pyrophosphorylase
Glucose-1-phosphate + UTP UDP-glucose + PPi
B. Synthesis of a primer to initiate glycogen synthesis

• A polypeptide of 332 amino acids called glycogenin

serves as a primer for glycogen synthesis
• Glycogenin is a self glucosylating enzyme that uses
UDP- glucose to link glucose to one of its own
tyrosine residue
• Glycogenin forms a chain of glucosyl residue on
itself by catalyzing the addition of seven more
glucosyl residue with α 1-4 linkage
• Glycosylated glycogenin then serves as a primer for
synthesis of glycogen by glycogen synthase.
C. Elongation of chain by glycogen synthase
• Glycogen synthase responsible for formation of
α1,4-glycosidic linkages.
• This enzyme transfers the glucose from UDP
glucose to the non-reducing end of glycogen to
form α-1,4 linkages
• Addition of glucose units to the glycogen primer
continues until the chain contains about eleven
glucose units
 D. Formation of branches
• When a growing chain is at least 11 glucose residues
long, branching enzyme transfers a part of the 1 → 4-
chain (at least six glucose residues) to a neighboring
chain to form a 1 → 6 linkage, establishing a branch
point.
• The two branches start growing again by addition of
glucose units by α-1,4-glycosidic bonds catalyzed by
glycogen Synthase
• When the branches contain about 11 glucose units,
branching enzyme acts again and creates more branches.
• In addition, the new branch point must be at least 4
residues away from a preexisting one
 .

• Branching is important because it increases the

solubility of glycogen
• Furthermore, branching creates a large number
of terminal residues, the sites of action of
glycogen phosphorylase and synthase. Thus,
branching increases the rate of glycogen
synthesis and degradation
 REGULATION
• The regulatory enzyme is GLYCOGEN
SYNTHASE which is regulated by
-allosteric mechanisms and
-covalent modification due to reversible
phosphorylation and dephosphorylation of
enzyme protein in response to hormone
action
REGULATION BY ALLOSTERIC MECHANISMS

• Glycogen Synthase is allosterically activated

by glucose-6-P.
• Thus Glycogen Synthase is active when high
blood glucose leads to elevated intracellular
glucose-6-P.
• It is useful to a cell to store glucose as
glycogen when the input to Glycolysis
(glucose-6-P), and the main product of
Glycolysis (ATP), are adequate.
REGULATION BY COVALENT MODIFICATION

• The hormones glucagon and epinephrine

activate G-protein coupled receptors to trigger
cAMP cascades.
• Both hormones are produced in response to
low blood sugar.
• Glucagon, which is synthesized by cells of the
pancreas, activates cAMP formation in liver.
• Muscle cells lack Glucagon receptors
• Epinephrine activates cAMP formation in
muscle
REGULATION BY COVALENT MODIFICATION

• Glycogen synthase exists in both

phosphorylated or nonphosphorylated states
• Active glycogen synthase a is
dephosphorylated and inactive glycogen
synthase b is phosphorylated
• The cAMP cascade results in phosphorylation
of a serine hydroxyl of Glycogen synthase,
which promotes transition to the inactive state.
REGULATION BY COVALENT MODIFICATION

• Phosphorylation of Glycogen Synthase

promotes the "b" (less active) conformation.
• The cAMP cascade thus inhibits glycogen
synthesis.
• Instead of being converted to glycogen,
glucose-1-P in liver may be converted to
glucose-6-P, and dephosphorylated for release
to the blood.
 ROLE OF INSULIN IN GLYCOGENESIS
• Insulin, produced in response to high blood
glucose, triggers a separate signal cascade that
leads to activation of Phosphoprotein Phosphatase.
• This phosphatase catalyzes removal of regulatory
phosphate residues from Glycogen Synthase
enzyme.
• Thus insulin antagonizes effects of the cAMP
cascade induced by glucagon & epinephrine.
• cAMP is hydrolyzed by phosphodiesterase, so
terminating hormone action; in liver insulin
increases the activity of phosphodiesterase.
 ..
INSULIN

+
Glc-6-PO4
+
.
GLYCOGENOLYSIS (DEGRADATION OF GLYCOGEN)

Glycogen is degraded by breaking α-1,4 and α-

1,6 glycosidic bonds
Glycogen degradation consists of following steps:
1. Action of GLYCOGEN PHOSPHORYLASE.
2. Action of GLUCAN TRANSFERASE
3. Action of DEBRANCHING ENZYME.
4. Action of PHOSPHOGLUCOMUTASE
5. Action of GLUCOSE-6-PHOSPHATASE (in LIVER
only)
. …..
 GLCOGEN
PHOSPHORYLASE
GLYCOGEN
MOLECULE

GLUCAN
TRANSFERASE

DEBRANCHING
ENZYME
PHOSPHORYLASE

PHOSPHOGLUCOMUTASE

GLUCOSE-6-PHOSPHATASE

GLYCOLYSIS
 1. ACTION OF GLYCOGEN PHOSPHORYLASE

• Rate regulating enzyme of glycogenolysis.

• Glycogen phosphorylase, cleaves its substrate by
the addition of orthophosphate (Pi) to yield glucose
-1-phosphate.
• The cleavage of a bond by the addition of
orthophosphate is referred to as phosphorolysis.
• Glycogen phosphorylase contain pyridoxal
phosphate as the coenzyme.
• Sequentially glucose-1-phosphate units are released
till about a four glucose residue long chain remains
at the branch point.
 2. ACTION OF GLUCAN TRANSFERASE

• Glucan transferase enzyme then transfers a

trisaccharide unit to a nearby branch.
• This exposes the branch point
i.e. α-1,6glycosidic bonds
 3. ACTION OF DEBRANCHING ENZYME .
• Debranching enzyme then splits the branch
point to release one more glucose unit as free
glucose and not as glu-1- phos.
• The remaining chain is then again acted upon
by phosphorylase
REMEMBER
Glucan transferase and Debranching enzyme
activities are associated with a single
bifunctional enzyme protein.
 4. ACTION OF PHOSPHOGLUCOMUTASE
• PHOSPHOGLUCOMUTASE
converts the glucose -1-
phosphate units into glucose
-6- phosphate in a reversible
reaction.
• Glucose 6-phosphate
derived from glycogen can
(1) be used as a fuel for
muscle (2) be converted into
free glucose in the liver and
subsequently released into
the blood; (3) be processed
by the pentose phosphate
pathway
 ACTION OF GLUCOSE-6-PHOSPHATASE (IN LIVER
ONLY)

• The liver contains a hydrolytic enzyme,

glucose-6-phosphatase, which cleaves the
phosphoryl group to form free glucose and
orthophosphate.
• This enzyme is also present in kidney.

Glu– 6-phosphate + H2O Glucose + Pi

The free glucose goes to blood and helps in
maintaining the blood glucose level.
 .
MUSCLE DOES NOT HAVE GLUCOSE -6- PHOSPHATASE

1. The end product in muscle is glucose -6-

phosphate and not free glucose because
muscles do not have enzyme glucose -6-
phosphate.
2. Muscles utilizes this glucose -6- phosphate to
obtain energy through glycolysis
DEGRADATION OF GLYCOGEN BY LYSOSOME

• Glycogen granules can also be engulfed by

lysosomes, where acid maltase catalyzes the
hydrolysis of glycogen to glucose.
• This may be especially important in glucose
homeostasis in neonates.
• Genetic lack of lysosomal acid maltase causes
type II glycogen storage disease (Pompe
disease,
• The lysosomal catabolism of glycogen is under
hormonal control.
 REGULATION
• Glycogen Synthase and Glycogen Phosphorylase
are reciprocally regulated, by allosteric effectors
and by phosphorylation
• The control of phosphorylase differs between
liver & muscle
• In the liver the role of glycogen is to provide free
glucose for export to maintain the blood
concentration of glucose
• In muscle the role of glycogen is to provide a
source of glucose 6-phosphate for glycolysis in
response to the need for ATP for muscle
contraction
 REGULATION

• The principal enzymes controlling glycogen

degradation—GLYCOGEN PHOSPHORYLASE are
regulated by three mechanisms:

1. Allosteric regulation
2. Hormonal regulation
3. Influence of calcium
 1. ALLOSTERIC REGULATION
• Adequate glucose and energy level inhibit
glycogenolysis
• Hence, Hepatic Phosphorylase is inhibited by glucose,
ATP, and glucose-6-phosphate.

•Muscle Phosphorylase is
inhibited by only ATP, and
glucose-6-phosphate and
not by free glucose.
•AMP is activator of it.
 2. HORMONAL REGULATION
• The enzyme phosphorylase Phosphorylase kinase is itself
occurs in two form both in regulated by phosphorylation
liver and muscle. and dephosphorylation and
• Phosphorylase a is active require Ca +2 binding for full
and phosphorylated form. activation
• Phosphorylase b is inactive
and dephosphorylated form.
• The two form are
interconvertible by enzyme
phosphorylase kinase and
protein phosphatase

FIG
 Phosphorylase Kinase
• Tetramer of four different subunits- α, β ,Υ and δ .
• The α and β subunits - phosphorylated by cAMP-
dependent protein kinase.
• The δ subunit is identical to the Ca2+- binding protein
calmodulin.
• The binding of Ca2+ activates the catalytic site of the
subunit Υ even while the enzyme is in the
dephosphorylated b state; the phosphorylated a form is
only fully activated in the presence of Ca2+.

Phosphorylase Kinase inactive

Phosphorylase Kinase- Ca++ partly active
P-Phosphorylase Kinase-Ca++ fully active
 cAMP .

Increasing the
concentration of cAMP
activates cAMPdependent
protein kinase, which
catalyzes the
phosphorylation of inactive
phosphorylase kinase b to
active phosphorylase
kinase a, which in turn,
phosphorylates
phosphorylase b to
phosphorylase a.
 .

• In the liver, cAMP is formed in response to

glucagon, which is secreted in response to
falling blood glucose;
• Muscle is insensitive to glucagon.
 .

• In muscle, the signal for increased cAMP

formation is the action of norepinephrine,
which is secreted in response to fear or fright,
when there is a need for increased
glycogenolysis to permit rapid muscle activity.
 .

FLIGHT/ FRIGHT
REACTION
CONTROL OF GLYCOGEN SYNTHASE IN MUSCLE.
 GLYCOGEN STORAGE DISEASES

A group of inherited disorders characterized by

deposition of an abnormal type or quantity of
glycogen in tissues, or failure to mobilize
glycogen.
 .
.
 Symptoms in addition to excess glycogen storage:
When a genetic defect affects mainly an isoform
of an enzyme expressed in liver, a common
symptom is hypoglycemia, relating to impaired
mobilization of glucose for release to the blood
during fasting.
When the defect is in muscle tissue, weakness &
difficulty with exercise result from inability to
increase glucose entry into Glycolysis during
exercise.
Additional symptoms depend on the particular
enzyme that is deficient.
 Von Gierke's disease

• It is transmitted by autosomal recessive trait.

• Enzyme deficiency: Glucose 6-phosphatase.
• The enzyme is absent in liver, kidney &
intestinal mucosa.
• Two types:
• Ia- Glucose 6-phosphatase enzyme is defective
• Ib- Endoplasmic reticulum glucose-6phosphate
transporter defective
• .
 Von Gierke's disease
Liver cells, intestinal mucosa & renal tubular epithelial
cells are loaded with glycogen, which is normal in
structure but metabolically not available
SYMPTOMS

Fasting hypoglycemia:
• Hypoglycemia, decreases insulin secretion, which in
turn inhibits protein synthesis causes stunted growth
(dwarfism).
Lactic acidemia:
• Glucose is not synthesized from lactate produced in
muscle & liver. Lactate level in blood increases & the
pH is lowered (acidosis).
 Von Gierke's disease
Hyperlipidemia:
• Fat is utilized as energy Hyperuricemia:
source, which leads to• Glucose 6-phosphate that
lipaemia, acidaemia & accumulates is diverted to
ketosis. HMP Shunt, leading to
• Excess of acetyl CoA increased synthesis of
resulting in increased ribose phosphates which
cholesterol levels, increase the cellular levels
produce xanthomas. of
• There is a blockade in phosphoribosylpyrophosph
ate & enhance the
gluconeogenesis. metabolism of purine
Ketosis due to FA oxidation nucleotides to uric acid
following hypoglycaemia (gouty arthritis).
Pi
.