Glycogen Metabolism

Copyright 1999-2007 by Joyce J. Diwan.

All rights reserved.
Biochemistry oI Metabolism
Glycogen is a polymer oI glucose residues linked by
-14) glycosidic bonds, mainly
-1) glycosidic bonds, at branch points.
Glycogen chains & branches are longer than shown.
Glucose is stored as glycogen predominantly in liver and
muscle cells.

H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
H
OH H
OH
CH
2
OH
H
O
H H H
O
O
H
OH H
OH
CH
2
H
H H
O
H
OH H
OH
CH
2
OH
H
OH
H H
O
O
H
OH H
OH
CH
2
OH
H
O
H
O
1 4

H
O
H
OH
H
OH
CH
2
OH
H
H H
O
H
OH H
OH
CH
2
OH
H
H
O
1
OH
3
4
5
2
glycogen
Glycogen Phosphorylase catalyzes phosphorolytic
cleavage oI the - glycosidic linkages oI glycogen,
releasing glucose-1-phosphate as reaction product.
glycogen
n residues
P
i

glycogen
n- residues
glucose--phosphate
%his phosphorolysis may be compared to hydrolysis:
Hydrolysis: ## ##
!hosphorolysis: R-O-R' + HO-PO
3
2-
R-OH + R'-O-PO
3
2-

glucose-1-phosphate
H
O
OH
H
OH H
OH
CH
2
OH
H
OPO
3
2
H
Glycogen catabolism
breakdown):
Pyridoxal phosphate
PLP), a derivative oI
vitamin B
6
, serves as
prosthetic group Ior
Glycogen !hosphorylase.
pyridoxal phosphate !!)

H
C
O
P
O
O
O
OH
CH
3
C
H O

H
2
!yridoxal phosphate !!) is held at the active site by a
$chiff base linkage, Iormed by reaction oI the aldehyde oI
!! with the 1-amino group oI a lysine residue.
In contrast to its role in other enzymes, the phosphate oI
!! is involved in acid/base catalysis by !hosphorylase.

H
C
O
P
O
O
O
O

CH
3
HC

H
2

CH
2
)
4
Enz
H

nzyme ys)-!! SchiII base

H
3

C COO

CH
2
CH
2
CH
2
CH
2
H
3
H

lysine
%he P
i
substrate binds
between the phosphate oI
!! and the glycosidic O
linking the terminal glucose
residue oI the glycogen.

H
C
O
P
O
O
O
O

CH
3
HC

H
2

CH
2
)
4
Enz
H

nzyme ys)-!! SchiII base

AIter the phosphate substrate donates

during cleavage
oI the glycosidic bond, it receives H

Irom the phosphate

moiety oI !!.
!! then takes back the H

as the phosphate O attacks C1

oI the cleaved glucose to yield glucose-1-phosphate.
A glucose analog-acetylglucosamine GlcNAc), is
adjacent to pyridoxal phosphate at the active site in the
crystal structure shown.
Glycogen
Phosphorylase:
a homodimeric
enzyme, subject to
allosteric control.
It transitions between
'relaxed active) &
'tense inhibited)
conIormations.
Diagramcomparing
relaxed and tense
conIormations.

!!
!!
GlcNAc
GlcNAc
inhibitor
Human iver
Glycogen !hosphorylase !DB 1M
"uestion: hy would an inhibitor oI Glycogen
!hosphorylase be a suitable treatment Ior diabetes?
A class oI drugs
developed Ior treating
the hyperglycemia oI
diabetes chloroindole-
carboxamides), inhibit
liver !hosphorylase
allosterically.
%hese inhibitors bind
at the dimer interIace,
stabilizing the inactive
tense) conIormation.

!!
!!
GlcNAc
GlcNAc
inhibitor
Human iver
Glycogen !hosphorylase !DB 1M
A glycogen storage site on the surIace oI the
!hosphorylase enzyme binds the glycogen particle.
Given the distance between storage & active sites,
!hosphorylase can cleave - linkages only to
within 4 residues oI an -6 branch point.
%his is called a "limit branch".
Explore the structure oI muscle Glycogen
!hosphorylase with Chime.
Debranching enzyme has 2 independent active sites,
consisting oI residues in diIIerent segments oI a single
polypeptide chain:
%he transferase oI the debranching enzyme
transIers 3 glucose residues Irom a 4-residue limit
branch to the end oI another branch, diminishing the
limit branch to a single glucose residue.
%he -6 glucosidase moiety oI the debranching
enzyme then catalyzes hydrolysis oI the -6
linkage, yielding Iree glucose. %his is a minor
Iraction oI glucose released Irom glycogen.
'iew an animation
%he major product oI glycogen breakdown is
glucose--phosphate, Irom !hosphorylase activity.
Phosphoglucomutase catalyzes the reversible reaction:
glucose--phosphate glucose-6-phosphate
A serine O at the active site donates & accepts !
i
.
%he bisphosphate is not released.
!hosphoglycerate Mutase has a similar mechanism, but
instead uses His Ior !
i
transIer.

glucose-1-phosphate glucose--phosphate
H
O
OH
H
OH H
OH
CH
2
OH
H
OPO
3
2
H H
O
OH
H
OH H
OH
CH
2
OPO
3
2
H
OH
H H
O
OH
H
OH H
OH
CH
2
OPO
3
2
H
OPO
3
2
H
Enzyme-Ser-OPO
3
2
Enzyme-Ser-OPO
3
2
Enzyme-Ser-OH
Glucose-6-phosphate may enter Glycolysis or mainly in
liver) be dephosphorylated Ior release to the blood.
iver Glucose-6-phosphatase catalyzes the Iollowing,
essential to the liver's role in maintaining blood glucose:
glucose-6-phosphate

O glucose P
i
Most other tissues lack this enzyme.

Glycogen Glucose
Hexokinase or Glucokinase
Glucose--!ase
Glucose-1-! Glucose--! Glucose !
i

Glycolysis
!athway
!yruvate
Glucose metabolism in liver.
&ridine diphosphate glucose &D!-glucose) is the
immediate precursor Ior glycogen synthesis.
As glucose residues are added to glycogen, &D!-glucose
is the substrate and &D! is released as a reaction product.
Nucleotide diphosphate sugars are precursors also Ior
synthesis oI other complex carbohydrates, including
oligosaccharide chains oI glycoproteins, etc.

O
O
OH OH
H H
H
CH
2
H
H

O
O
O P
O
O

P
O
O

H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
&D!-glucose
Glycogen
synthesis

O
O
OH OH
H H
H
CH
2
H
H

O
O
O P
O
O

P
O
O

H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
O

P
O
O

H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
O
O
OH OH
H H
H
CH
2
H
H

O
O
O P
O
O

P
O
O

O P

O
O
O

PP
i

&D!-glucose
glucose-1-phosphate
&%!
&D!-Glucose !yrophosphorylase
&DP-glucose is Iormed Irom glucose-1-phosphate:
glucose--phosphate &%P &DP-glucose PP
i
PP
i

O P
i
Overall:
glucose--phosphate &%P &DP-glucose P
i
Spontaneous hydrolysis oI the =! bond in !!
i
!=!)
drives the overall reaction.
Cleavage oI !!
i
is the only energy cost Ior glycogen
synthesis one =! bond per glucose residue).
Glycogenin initiates glycogen synthesis.
Glycogenin is an enzyme that catalyzes attachment oI a
glucose molecule to one oI its own tyrosine residues.
Glycogenin is a dimer, and evidence indicates that the
2 copies oI the enzyme glucosylate one another.

%yr active site

active site %yr
Glycogenin dimer
A glycosidic bond is Iormed between the anomeric C1 oI
the glucose moiety derived Irom &D!-glucose and the
hydroxyl oxygen oI a tyrosine side-chain oI Glycogenin.
&D! is released as a product.
H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
H
OH H
OH
CH
2
OH
H
O
H H
C
CH
H
C
H
2
O
O
H
O
OH
H
OH H
OH
CH
2
OH
H
H
C
CH
H
C
H
2
O
O
1
5
4
3
2
6
H
O
OH
H
OH H
OH
CH
2
OH
H
H
O
1
5
4
3
2
6
P O P O Uridine
O
O

O
O

C
CH
H
C
H
2
HO
O
tyrosine residue
oI Glycogenin

O-linked
glucose
residue
&D!
&D!-glucose
Glycogenin then catalyzes glucosylation at C4 oI the
attached glucose &D!-glucose again the donor), to yield an
O-linked disaccharide with -1F4) glycosidic linkage.
%his is repeated until a short linear glucose polymer with
-1F4) glycosidic linkages is built up on Glycogenin.
H
O
OH
H
OH H
OH
CH
2
OH
H
O
H
H
OH H
OH
CH
2
OH
H
O
H H
C
CH
H
C
H
2
O
O
H
O
OH
H
OH H
OH
CH
2
OH
H
H
C
CH
H
C
H
2
O
O
1
5
4
3
2
6
O O

&D!-glucose
O-linked
glucose
residue
-1 4)
linkage
&D!
&D!
nswer: Most oI the Glycogenin is Iound associated
with glycogen particles branched glycogen chains)
in the cytoplasm.
Glycogen $ynthase then catalyzes elongation oI
glycogen chains initiated by Glycogenin.
"uestion: here would you expect to Iind
Glycogenin within a cell?
Glycogen $ynthase catalyzes transIer oI the glucose
moiety oI &D!-glucose to the hydroxyl at C4 oI the
terminal residue oI a glycogen chain to Iorm an
-F glycosidic linkage:
glycogen
n residues
&DP-glucose
glycogen
n residues
&DP
A branching enzyme transIers a segment Irom the end oI
a glycogen chain to the C hydroxyl oI a glucose residue
oI glycogen to yield a branch with an -F6 linkage.
Both synthesis & breakdown oI glycogen are spontaneous.
II both pathways were active simultaneously in a cell,
there would be a "futile cycle" with cleavage oI one ~P
bond per cycle in Iorming &D!-glucose).
%o prevent such a Iutile cycle, Glycogen Synthase and
Glycogen !hosphorylase are reciprocally regulated, by
allosteric eIIectors and by phosphorylation.
Glycogen Synthesis

&%! &D! 2 !
i


glycogen
n)
glucose-1-! glycogen
n 1)


Glycogen !hosphorylase !
i


Glycogen Phosphorylase in muscle is subject to allosteric
regulation by AM!, A%!, and glucose--phosphate.
A separate isozyme oI !hosphorylase expressed in liver is
less sensitive to these allosteric controls.
P present signiIicantly when A%! is depleted)
activates !hosphorylase, promoting the relaxed
conIormation.
%P & glucose-6-phosphate, which both have
binding sites that overlap that oI AM!, inhibit
!hosphorylase, promoting the tense conIormation.
%hus glycogen breakdown is inhibited when A%! and
glucose--phosphate are plentiIul.
Glycogen $ynthase is allosterically activated by
glucose-6-P opposite oI eIIect on !hosphorylase).
%hus Glycogen Synthase is active when high blood
glucose leads to elevated intracellular glucose--!.
It is useIul to a cell to store glucose as glycogen when the
input to Glycolysis glucose--!), and the main product
oI Glycolysis A%!), are adequate.

Glycogen Glucose
Hexokinase or Glucokinase
Glucose--!ase
Glucose-1-! Glucose--! Glucose !
i

Glycolysis
!athway
!yruvate
Glucose metabolism in liver.
#egulation by covalent modification
phosphorylation):
%he hormones glucagon and epinephrine activate
G-protein coupled receptors to trigger cP cascades.
Both hormones are produced in response to low
blood sugar.
Glucagon, which is synthesized by --cells oI the
pancreas, activates cAM! Iormation in liver.
pinephrine activates cAM! Iormation in muscle.
%he cAM! cascade results in phosphorylation oI a serine
hydroxyl oI Glycogen !hosphorylase, which promotes
transition to the active relaxed) state.
%he phosphorylated enzyme is less sensitive to allosteric
inhibitors.
%hus, even iI cellular A%! & glucose--phosphate are
high, !hosphorylase will be active.
%he glucose-1-phosphate produced Irom glycogen in liver
may be converted to Iree glucose Ior release to the blood.
ith this hormone-activated regulation, the needs oI the
organism take precedence over needs oI the cell.
Commonly used terminology:
"a" is the Iorm oI the enzyme that tends to be active,
and independent oI allosteric regulators in the case
oI Glycogen !hosphorylase, when phosphorylated).
"b" is the Iorm oI the enzyme that is dependent on
local allosteric controls in the case oI Glycogen
!hosphorylase when dephosphorylated).
Signal
cascade by
which
Glycogen
!hosphorylase
is activated.
Hormone epinephrine or glucagon)
via G !rotein G
-
-G%!)
Adenylate cyclase Adenylate cyclase
inactive) active)
catalysis

A%! cyclic AM! !!
i

Activation !hosphodiesterase
AM!
!rotein kinase A !rotein kinase A
inactive) active)
A%!
AD!
!hosphorylase kinase !hosphorylase kinase !)
b-inactive) a-active)
!hosphatase A%!
!
i
AD!

!hosphorylase !hosphorylase !)
b-allosteric) a-active)
!hosphatase
!
i

%he cP cascade induced in liver by glucagon or
epinephrine has the opposite effect on glycogen
synthesis.
Glycogen $ynthase is phosphorylated by !rotein
Kinase A as well as by !hosphorylase Kinase.
Phosphorylation oI Glycogen Synthase promotes the
"b" less active) conIormation.
%he cAM! cascade thus inhibits glycogen synthesis.
Instead oI being converted to glycogen, glucose-1-!
in liver may be converted to glucose--!, and
dephosphorylated Ior release to the blood.
High cytosolic glucose--phosphate, which would result
when blood glucose is high, turns oII the signal with
regard to glycogen synthesis.
%he conIormation oI Glycogen Synthase induced by the
allosteric activator glucose--phosphate is susceptible to
dephosphorylation by !rotein !hosphatase.

Glycogen Glucose
Hexokinase or Glucokinase
Glucose--!ase
Glucose-1-! Glucose--! Glucose !
i

Glycolysis
!athway
!yruvate
Glucose metabolism in liver.
nsulin, produced in response to high blood glucose,
triggers a separate signal cascade that leads to
activation of Phosphoprotein Phosphatase.
%his phosphatase catalyzes removal oI regulatory
phosphate residues Irom !hosphorylase, !hosphorylase
Kinase, & Glycogen Synthase enzymes.
%hus insulin antagonizes eIIects oI the cAM! cascade
induced by glucagon & epinephrine.
a

also regulates glycogen breakdown in muscle.

During activation oI contraction in skeletal muscle, a

is released Irom the sarcoplasmic reticulum to promote

actin/myosin interactions.
%he released a

also activates Phosphorylase Kinase,

which in muscle includes calmodulin as its 0 subunit.
!hosphorylase Kinase is partly activated by binding oI
a

to this subunit.
!hosphorylase Kinase inactive

!hosphorylase Kinase-Ca

partly active


!-!hosphorylase Kinase-Ca

Iully active
Phosphorylation oI the enzyme, via a cAM! cascade
induced by epinephrine, results in Iurther activation.
%hese regulatory processes ensure release oI phosphorylated
glucose Irom glycogen, Ior entry into Glycolysis to provide
%P needed Ior muscle contraction.
During extended exercise, as glycogen stores become
depleted, muscle cells rely more on glucose uptake Irom
the blood, and on Iatty acid catabolism as a source oI A%!.
!hosphorylase Kinase inactive

!hosphorylase Kinase-Ca

partly active


!-!hosphorylase Kinase-Ca

Iully active
A genetic defect in the isoIorm oI an enzyme expressed in
liver causes the Iollowing symptoms:
fter eating a O meal, elevated blood levels oI
glucose, lactate, & lipids.
During fasting, low blood glucose & high ketone bodies.
Which liver enzyme is deIective?
Explain $ymptoms:
fter eating, blood glucose is high because liver cannot
store it as glycogen. Some excess glucose is processed
via Glycolysis to produce lactate & Iatty acid precursors.
During fasting, glucose is low because the liver lacks
glycogen stores Ior generation oI glucose.
Ketone bodies are produced as an alternative fuel.
Glycogen $ynthase
"uestion: How would you nutritionally treat
deIiciency oI liver Glycogen Synthase?
requent meals oI complex carbohydrates
avoiding simple sugars that would lead to a rapid
rise in blood glucose)
Meals high in protein to provide substrates Ior
gluconeogenesis.
Glycogen $torage
Diseases are genetic
enzyme deIiciencies
associated with excessive
glycogen accumulation
within cells.
Some enzymes whose
deIiciency leads to
glycogen accumulation
are part oI the inter-
connected pathways
shown here.
glycogen

glucose-1-!
Glucose--!hosphatase
glucose--! glucose !
i


Iructose--!
!hosphoIructokinase
Iructose-1,-bis!

Glycolysis continued
$ymptoms in addition to excess glycogen storage:
hen a genetic deIect aIIects mainly an isoIorm oI an
enzyme expressed in liver, a common symptom is
hypoglycemia, relating to impaired mobilization oI
glucose Ior release to the blood during Iasting.
hen the deIect is in muscle tissue, weakness &
difficulty with exercise result Irom inability to
increase glucose entry into Glycolysis during exercise.
Additional symptoms depend on the particular
enzyme that is deIicient.
Glycogen $torage Disease
$ymptoms, in addition to
glycogen accumulation
%ype , liver deIiciency oI
Glucose-6-phosphatase von
Gierke's disease)
hypoglycemia low blood
glucose) when Iasting, liver
enlargement.
%ype ', deIiciency oI
branching enzyme in various
organs, including liver
Andersen's disease)
liver dysfunction and early
death.
%ype ', muscle deIiciency oI
Glycogen Phosphorylase
McArdle's disease)
muscle cramps with exercise.
%ype ', muscle deIiciency oI
Phosphofructokinase.
inability to exercise.