GLYCOGEN METABOLISM

Learning objectives:

Describe composition and glycosidic bonds in

glycogen

Describe the biochemical pathway of glycogen

synthesis

Describe the biochemical pathway of

glycogenolysis

Discuss regulation of glycogen metabolism

 Glycogen

α-1,4

α-1,4
CH
H 2O
H
O H O α-1,6
OH O
CH2OH CH2OH CH2 CH2OH CH2OH CH2OH CH2 CH2OH
H CH
O O O H O HH H 2 O O H H O H O H O H
H H H H H HH H H H H
H H H H H H H H H
….. O
OH H
O
OH H
O
OH H
O
OH
OH
O
H OH
O
OH O
H
H
O
OH H
O
OH H
O
OH H ….
CH
H H α-1,42 O H α-1,4 α-1,4
H
H OH H OH H OH H OH H OH H OH H OH H OH
O H
O O
OH H
H CH
H H 2O
H H

Glycogen is a branched homopolysaccharide composed

OH
O
OHof α-D-glucose
H
O units
H
bound by α-1,4 and (at branch points) α-1,6 glycosidicH bonds.H

OH

On average, there are branches for every 8-10 glycosyl residues.

 Glycogen

α-1,4

α-1,4
CH
H 2O
H
O H O α-1,6
OH O
CH2OH CH2OH CH2 CH2OH CH2OH CH2OH CH2 CH2OH
H CH
O O O H O HH H 2 O O H H O H O H O H
H H H H H HH H H H H
H H H H H H H H H
….. O
OH H
O
OH H
O
OH H
O
OH
OH
O
H OH
O
OH O
H
H
O
OH H
O
OH H
O
OH H ….
CH
H H α-1,42 O H α-1,4 α-1,4
H
H OH H OH H OH H OH H OH H OH H OH H OH
O H
O O
OH H
H CH
H H 2O
H
A single molecule can have a molecular mass of upO to H108 Da
O
with H

OH
more than 500,000 glucosyl residues.
OH
H
H H

O
Glycogen forms intracellular glycogen granules in the cytoplasm.
H
Electron micrograph of a section of a liver cell showing glycogen deposits
as accumulations of electron dense particles (arrows).
Glycosyl residue attached by
an α-1,6 glycosidic bond

Glycosyl residue at a
non-reducing end

Glycosyl units are attached

and mobilized from the
reducing ends
Glycogen is an intracellular storage form of readily available glucose

Main stores of glycogen in the human body:

Liver - Approximately 100 g or 10% of the fresh weight
Muscle - Approximately 400 g or 1-2% of the fresh weight

Most other cells have small amounts of glycogen stored

 LIVER MUSCLE

Glycogen
Glycogen
Glucose 6-P
Glucose 6-P
G6Pase

Glucose
GLYCOLYSIS

Blood glucose
 Sources of blood glucose after a meal
mM glucose
8

Meal

Glycogen

Gluconeogenesis

8 16 24 2 7 30

Hours Days
 Glycogen synthesis
Glycogenesis

Glycogen is synthesized from molecules of α-D-glucose.

Synthesis occurs in the cytosol

Synthesis requires energy

ATP for phosphorylation of glucose
UTP for generating an activated form of glucose: UDP-glucose
 Glycogen synthesis - Glycogenesis
ATP Glucose
Hexokinase/Glucokinase
ADP
Glucose 6-phosphate
Phosphoglucomutase

UTP Glucose 1-phosphate

Pyrophosphatase UDP-glucose pyrophosphorylase
2 Pi + H2O PPi
UDP-glucose
Glycogenn
Glycogen synthase

Glycogenn+1
Branching enzyme
 CH2OH Glucokinase CH2OPO32-

H O H Hexokinase O
H H
H + ATP H + ADP
OH H OH H
OH OH OH OH

H OH H OH
Glucose Glucose 6-phosphate

Same reaction, same enzymes, and same regulation as in glycolysis

Irreversible

Hexokinase

- Glucose 6-phosphate (low phosphofructokinase activity)

Glucokinase

+ High blood glucose (release from GKRP, High Km)

+ Insulin stimulates gene transcription (only in liver)

Phosphoglucomutase
OPO32-
Ser CH2OPO32-

H O H
H
CH2OPO32-
OH H
H O H OH OH
OH H
Ser H OH
OH H
Glucose 6-phosphate
OH OPO32-
CH2OH
H OH
H O H
Glucose 1,6-bisphosphate H
OH H
OH OPO32-

OPO32- H OH
Ser
Glucose 1-phosphate
 CH2OH

H O H O O O
H
+ O- – P - O – P – O – P – O - uridine
OH H
OH OPO32-
O- O- O-
H OH
Glucose 1-phosphate UTP

UDP-glucose pyrophosphorylase
CH2OH

H O H
H O O O O
OH H
OH O – P – O – P – O - uridine + O- – P – O – P – O -

H OH O- O- O- O-

UDP-glucose Pyrophosphate (PPi)

 O O Pyrophosphatase

O- – P – O – P – O - + H2O 2 Pi

O- O-
NB: Irreversible reaction
Pyrophosphate (PPi)

Glucose 1-phosphate + UTP UDP-glucose + PP i

PPi + H2O 2 Pi

Glucose 1-phosphate + UTP + H2O UDP-glucose + 2 P i

The irreversible hydrolysis of pyrophosphate drives the synthesis of

UDP-glucose
 CH2OH CH2OH
H O H O
H H
H O O H
OH H OH H
OH O – P – O – P – O - uridine + O-R
HO
α-1,4
H OH O- O- H OH
UDP-glucose Glycogen (n residues)

Glycogen synthase

CH2 CH2OH
O H O H
H H
O O
H H
O- – P – O – P – O - uridine + OH H OH H
O O-R
HO
α-1,4 α-1,4
O- O-
H OH H OH
UDP Glycogen (n+1 residues)
 Priming of glycogen synthesis

Glycogen synthase can NOT add glucosyl residues to free glucose

or to oligosaccharides of less than 8 glucosyl residues

Priming is catalyzed by the protein GLYCOGENIN

The first glucosyl residue is attached in an O-glycosidic linkage

to the hydroxyl group of tyrosine of Glycogenin itself

7 additional residues are attached by glycogenin

Glycogenin remains attached to the reducing end of the glycogen

molecule
 HO Tyr
8 UDP-glucose +

Glycogenin

Glycogenin

O Tyr

Glycogenin
 …

Non-reducing end Cleaveage of α-1,4 bond

“Branching enzyme”
Amylo-α(1,4) → α(1,6)-transglucosidase

α-1,6
Non-reducing ends bond

…
 Stoichiometry

Glucose + ATP + UTP + H2O + Glycogenn →

Glycogenn+1 + ADP + UDP + 2 Pi

 Degradation of glycogen
Glycogenolysis

Occurs in cytoplasm

Major product is glucose 1-phosphate from breaking α-1,4 bonds

Minor product is glucose from breaking α-1,6 bonds

Glucose 1-phosphate : Glucose ≈ 10:1

 Glycogen synthesis - Glycogenesis
Pi Glycogenn
Glycogen phosphorylase

Glucose 1-phosphate Glycogenn-1

Phosphoglucomutase

…
Glucose 6-phosphate
H2O
G6Pase
Pi “Debranching
Glucose H2O
enzyme”
Glycogen with branch

Glycolysis
 CH2OH CH2 CH2OH

O H O H O H O H
H H
+ H H H
O- – P – OH
OH H OH H OH H
HO O O O-R
O-
α-1,4 α-1,4
H OH H OH H OH
Phosphate Glycogen with n residues

Glycogen phosphorylase

CH2OH CH2 CH2OH

O O H O H
H H H H
H H H
OH H + OH H OH H
O O-R
OH OPO32- HO
α-1,4
H OH H OH H OH

Glucose 1-phosphate Glycogen with n-1 residues

 H N Lys
C
2-
O3PO-CH2 OH

CH3
N+
H

Pyridoxal phosphate is a coenzyme for the phosphorylase

reaction.

Pyridoxal phosphate is bound to a nitrogen of a lysyl

residue of glycogen phosphorylase

The phosphate of pyridoxal phosphate exchanges protons with

the phosphate reactant, which allows the reactant to donate a
proton to the oxygen atom on carbon 4.
 CH2OH
Phosphoglucomutase O
OPO32- H H
Ser
H
OH H
OH OPO32-
CH2OPO32-
O H OH
H H
OH H Glucose 1-phosphate
Ser
OH H
OH OPO32-

H OH
CH2OPO32-
Glucose 1,6-bisphosphate
H O H
H
OH H
OPO32- OH OH
Ser
H OH
Glucose 6-phosphate
 Glucose-6-
CH2OPO 2- CH2OH
3
phosphatase
O O
H H (G6Pase) H H
H H
+ H2O OH H
+ Pi
OH H
OH OH OH OH

H OH H OH
Glucose 6-phosphate Glucose

Same reaction as in gluconeogenesis

Occurs in endoplasmic reticulum and involves a
glucose 6-phosphatase transporter and a catalytic subunit

The catalytic subunit is regulated at the level of transcription

 Glycogen phosphorylase stops when 4 glucosyl units
remain on each chain from a branch point
a’
b’
c’
α-1,6
d’
bond

a b c d e …
Oligo-α(1,4)→α(1,4)-glucan
transferase
(debranching enzyme) d’ α-1,6
bond
a’ b’ c’ a b c d e …

Amylo-α(1,6)-glucosidase H2O
(debranching enzyme)

d’
+ a’ b’ c’ a b c d e …
Glucose
 Approximate Stoichiometry

Glycogenn+11 + 10 Pi + H2O →

Glycogenn + 10 Glucose 6-phosphate + Glucose

 LIVER MUSCLE

Glycogen
Glycogen
Glucose 6-P
Glucose 6-P
G6Pase

Glucose
GLYCOLYSIS

Blood glucose
 Regulation of glycogen metabolism

Skeletal muscle
Glycogen must be broken down to provide ATP for
contraction, when the muscle is rapidly contracting,
or in anticipation of contractions in stress situations
like fear or excitement.

In rapidly contracting muscle: Low [ATP], High [AMP]

High [Ca++]

Stress: High [Epinephrine]

Glycogen stores are replenished when muscles are

resting.

Resting state: Low [AMP], High [ATP]

 Hormonal regulation of metabolism
Hormone Type Secreted by Secreted in response to

Insulin Protein Pancreatic beta cells High blood [glucose]

Glucagon Polypeptide Pancreatic alpha cells Low blood [glucose]

Epinephrine Catecholamine Adrenal medulla Stress

(adrenalin) Nervous system Low blood [glucose]

Glucocorticoids Steroid hormone Adrenal cortex Stress

Low blood [glucose]

Glucagon is the most important hormone signaling

low blood glucose concentration, while epinephrine and
glucocorticoids play secondary roles.
 Regulation of glycogen metabolism

Liver
Glycogen must be broken down to provide glucose
for maintaining blood glucose in fasting or for providing
additional glucose for skeletal muscles in stress situations.

Fasting: High [Glucagon]

Stress: High [Epinephrine]

Glycogen stores must be replenished in the fed state

Fed state: High [Insulin]

High [Glucose]
Muscle

Glycogen Glycogen

Glucose 6-phosphate Glucose 6-phosphate

Rapidly contracting state Resting state and with

Stress abundant energy

Liver
Glycogen Glycogen

Glucose 6-phosphate Glucose 6-phosphate

Fasting state Fed state

Stress
Key regulatory enzyme of glycogen breakdown:
Glycogen phosphorylase

Key regulatory enzyme of glycogen synthesis:

Glycogen synthase
Glycogen phosphorylase is a dimer of identical subunits.
Glycogen phosphorylase can exist in an active R (relaxed)
and an inactive T (tense) state.

In the T state, the catalytic site is partly blocked

Red: active site

Yellow:
Glycogen binding site

Red site:
Allosteric site for
AMP binding

Blue/green sites:
Phosphorylation sites
Allosteric regulation of glycogen phosphorylase
Regulation by energy state.

+ AMP (binding favors the active R state)

- ATP (binding favors the inactive T state)

Regulation by feedback inhibition.

- Glucose 6-phosphate (G6P)
G6P concentration increases when G6P is generated
faster than it can be further metabolized, e.g. by glycolysis

Regulation by high blood glucose

- Glucose (Only liver glycogen phosphorylase)

In the fed state with a high blood glucose concentration,
there is no need for the liver to secrete glucose
Regulation of glycogen phosphorylase by phosphorylation

Phosphorylase
kinase
ATP ADP

P
Glycogen Glycogen
phosphorylase b phosphorylase a

P
Inactive Active
T state Pi H2O
R state

Protein phosphatase 1
(PP1)

Phosphorylation occurs in the fasted or stressed state

Dephosphorylation is stimulated in the fed state
Phosphorylase kinase is regulated by phosphorylation and Ca ++ binding
One subunit is the Ca++ -binding calmodulin

Ca++ Ca++ PKA

Ca++
PP1
P P
Ca++ Ca++

Ca++ Ca++

P P

Ca++ Ca++
P P
PKA
Ca++

PP1 P P

Inactive
Inactive Partly active
Partly active Fully active
Fully active

Phosphorylation occurs in the fasted or stressed state.

Dephosphorylation is stimulated in the fed state.
Ca++ binding occurs when the [Ca++] is high, e.g. during rapid muscle contractions
Glucagon Epinephrine
(low blood glucose) (stress, fear)
Adenylyl cyclase

Cell
Glucagon receptor
(liver) + + Epinephrine receptor
(muscle and liver) membrane
ATP cAMP

+
Inactive Active
Protein kinase A Protein kinase A (PKA)

ATP ADP

Inactive Active P
Phosphorylase kinase Phosphorylase kinase

ATP ADP

Inactive glycogen Active glycogen

phosphorylase b phosphorylase a

Glycogenn Glycogenn-1

Pi Glucose 1-phosphate
 cAMP

Adenylyl cyclase
ATP cAMP + PPi

H2O
Phosphodiesterase

AMP
 Glucagon receptors and epinephrine receptors are
G-protein-coupled receptor
GDP

Adenylyl
GDP
cyclase

Receptor
beta and gamma subunit
of G-protein

alpha subunit
of G-protein
GTP

When hormone is no longer present, intrinsic GTP hydrolase activity of the G-protein alpha subunit
hydrolyzes GTP to GDP, the alpha subunit re-associates with the beta and gamma subunits, and
stimulation of adenylyl cyclase ends. cAMP is converted to AMP by phosphodiesterase.
Thus, in the absence of hormone, the cAMP concentration rapidly falls.
 Insulin α α α α α α

β β β β β β
P P P P
Insulin receptor
It functions as a P P P P
tyrosine kinase Autophosphorylation
when insulin P
is bound

Insulin receptor substrate

Activation Activation of
of protein multiple signaling
phosphatases pathways
Activation
of protein
kinases

In general, the protein kinases activated by insulin have opposite biological effects
from those activated by glucagon

In general, the protein phosphatases activated by insulin dephosphorylate proteins

that are phosphorylated by glucagon-stimulated protein kinases, such as PKA
 Regulation of glycogen synthase

Regulation by feed-forward mechanism.

+ Glucose 6-phosphate (G6P)
G6P concentration increases at high glucose concentrations
when G6P is generated faster than it can be further metabolized

NB: Reciprocal regulation of glycogen synthase

and glycogen phosphorylase by glucose 6-phosphate
Regulation of glycogen synthase by phosphorylation

PKA and Glycogen synthase kinase

ATP ADP

P
Active Inactive
Pi H2O

Protein phosphatase 1
(PP1)

Phosphorylation occurs in the fasted or stressed state

Dephosphorylation is stimulated in the fed state
Reciprocal regulation of glycogen phosphorylase and
glycogen synthase by phosphorylation
Fasting/stress
(glucagon/epinephrine)
+
ATP ADP

P P

Phosphorylase
kinase
PKA

Pi H2O P P

Active

ATP ADP
ATP ADP
P
P
Glycogen Glycogen
phosphorylase synthase
P
Inactive Active P
T state Pi R state Active Inactive
H2O
Pi H2O

PP1 + Fed state (insulin)

And it is even more complex..
Scaffolding proteins of different subtypes in liver and
muscle can bind the glycogen particle, PP1,
glycogen phosphorylase, and glycogen synthase

Binding brings participants of glycogen metabolism

together.

Regulation of PP1 is itself complex with various

inhibitors responding to the metabolic state of
the organism.