Fundamentals of

Biochemistry
Fourth Edition

Donald Voet Judith G. Voet

Charlotte W. Pratt

Chapter 16
Glycogen Metabolism and Gluconeogenesis

Copyright 2013 by John Wiley & Sons, Inc. All rights reserved.
 Storage Polysaccharides-Glycogen
Glycogen is a multibranched polysaccharide of glucose that serves as a form of energy
storage in animals and fungi. Glycogen is present in all cells but is most prevalent in
skeletal muscle and in liver.
The primary structure of glycogen resembles that of amylopectin, but glycogen is more
highly branched permitting the rapid mobilization of glucose in times of metabolic need.
In the cell, glycogen is degraded for metabolic use by glycogen phosphorylase and
glycogen debranching enzyme.

Glycogen granules in a liver cell

Glycogen (in animals, fungi, and bacteria) and starch (in plants) can function to stockpile glucose
for later metabolic use. In animals, a constant supply of glucose is essential for tissues such as the
brain and red blood cells, which depend almost entirely on glucose as an energy source (other
tissues can also oxidize fatty acids for energy; Section 20-2). The mobilization of glucose from
glycogen stores, primarily in the liver, provides a constant supply of glucose (~5 mM in blood) to
all tissues. When glucose is plentiful, such as immediately after a meal, glycogen synthesis
accelerates. Yet the liver's capacity to store glycogen is sufficient to supply the brain with glucose
for about half a day. Under fasting conditions, most of the body's glucose needs are met by
gluconeogenesis (literally, new glucose synthesis) from noncarbohydrate precursors such as amino
acids. Not surprisingly, the regulation of glucose synthesis, storage, mobilization, and
catabolism by glycolysis (Section 15-2) or the pentose phosphate pathway (Section 15-6) is
elaborate and is sensitive to the immediate and long-term energy needs of the organism.

The importance of glycogen for glucose storage is plainly illustrated by the effects of deficiencies
of the enzymes that release stored glucose. McArdle's disease, for example, is an inherited
condition whose major symptom is painful muscle cramps on exertion. The muscles in afflicted
individuals lack the enzyme required for glycogen breakdown to yield glucose. Although glycogen
is synthesized normally, it cannot supply fuel for glycolysis to keep up with the demand forATP.
 The metabolic uses of glucose

Glucose-6-phosphate (G6P) is a key branch point and has several possible fates: It can be used to
synthesize glycogen; it can be catabolized via glycolysis to yield ATP and carbon atoms (as acetyl-
CoA) that are further oxidized by the citric acid cycle; and it can be shunted through the pentose
phosphate pathway to generate NADPH and/or ribose-5-phosphate. In the liver and kidney, G6P
can be converted to glucose for export to other tissues via the bloodstream.
 Glycogen Breakdown

Glycogen granules are especially prominent in the cells that make the greatest use of
glycogen: muscle (up to 12% glycogen by weight) and liver cells (up to 10% glycogen
by weight.

Glucose units are mobilized by their sequential removal from the nonreducing ends of
glycogen (the ends lacking a C1-OH group). Whereas glycogen has only one reducing end,
there is a nonreducing end on every branch. Glycogen's highly branched structure therefore
permits rapid glucose mobilization through the simultaneous release of the glucose units at
the end of every branch.
Glycogen Phosphorylase Degrades Glycogen to Glucose-1-Phosphate

Phosphorylase covalently binds the

cofactor pyridoxal--5--phosphate which
is a vitamin B6 derivative. The
phosphorylase reaction mechanism which
shows how PLP's phosphate group
functions as a general acidbase catalyst.
The phosphorylation of Ser 14 promotes phosphorylase's T (inactive) R (active) conformational
change a manner that resembles allosteric control.

The T-state enzyme is inactive

because it has a malformed active
site and a surface loop that blocks
substrate access to its binding site.

The conformation of phosphorylase b is

allosterically controlled by the effectors AMP,
ATP, and G6P and is mostly in the T state
under physiological conditions. In contrast,
the phosphorylated form of the enzyme,
phosphorylase a, is unresponsive to these
effectors and is mostly in the R state unless
there is a high level of glucose. Thus, under
usual physiological conditions, the enzymatic
activity of glycogen phosphorylase is largely
determined by its rates of phosphorylation
and dephosphorylation.
 Glycogen Debranching Enzyme
Phosphorolysis proceeds along a glycogen branch until it approaches to within 4 or 5
residues of an (16) branch point, leaving a limit branch. Glycogen debranching
enzyme acts as an (14) transglycosylase (glycosyltransferase) by transferring an
(14)-linked trisaccharide unit from a limit branch of glycogen to the nonreducing end
of another branch.
This reaction forms a new (14) linkage with 3 more
units available for phosphorylase-catalyzed
phosphorolysis. The (16) bond linking the
remaining glycosyl residue in the branch to the main
chain is hydrolyzed (not phosphorylyzed) by the same
debranching enzyme to yield glucose and debranched
glycogen.About 10% of the residues in glycogen (those
at the branch points) are therefore converted to glucose
rather than G1P. Debranching enzyme has separate
active sites for the transferase and the (16)-
glucosidase reactions which improves the efficiency of
the debranching process.
 Phosphoglucomutase Interconverts Glucose-1-Phosphate and Glucose-6-Phosphate

Glucose-6-Phosphatase Generates Glucose in the Liver.

The G6P produced by glycogen breakdown can continue along the glycolytic
pathway or the pentose phosphate pathway. In the liver, G6P is also made
available for use by other tissues. Because G6P cannot pass through the cell
membrane, it is first hydrolyzed by glucose-6-phosphatase (G6Pase). G6Pase
resides in the endoplasmic reticulum (ER) membrane. Consequently G6P must
be imported into the ER by a G6P translocase before it can be hydrolyzed. The
resulting glucose and Pi are then returned to the cytosol via specific transport
proteins. A defect in any of the components of this G6P hydrolysis system results
in type I glycogen storage disease. Glucose leaves the liver cell via a specific
glucose transporter named GLUT2 and is carried by the blood to other tissues.
Muscle and other tissues lack G6Pase and therefore retain their G6P.
 Glycogen Synthesis
Glycogen synthesis and breakdown occur by separate pathways. The three enzymes that participate in
glycogen synthesis are UDPglucose pyrophosphorylase, glycogen synthase, and glycogen branching
enzyme.

The separation of synthetic and degradative pathways for glycogen was recognized by studying
McArdle's disease
UDPGlucose Pyrophosphorylase Activates Glucosyl Units

The reaction catalyzed by UDPglucose pyrophosphorylase

 Glycogen Synthase Extends Glycogen Chains

Glycogenin Primes Glycogen Synthesis. Glycogen synthase cannot simply link together two glucose
residues; it can only extend an already existing (14)-linked glucan chain. How, then, is glycogen
synthesis initiated? In the first step of this process, a 349-residue protein named glycogenin, acting as a
glycosyltransferase, attaches a glucose residue donated by UDPG to the OH group of its Tyr 194.
Glycogenin then extends the glucose chain by up to seven additional UDPG-donated glucose residues
to form a glycogen primer. Only at this point does glycogen synthase commence glycogen synthesis
by extending the primer. Analysis of glycogen granules suggests that each glycogen molecule is
associated with only one molecule each of glycogenin and glycogen synthase.
 Glycogen Branching Enzyme Transfers Seven-Residue Glycogen Segments

Branching to form glycogen is accomplished by branching enzyme. A branch is created

by transferring a 7-residue segment from the end of a chain to the C6-OH group of a
glucose residue on the same or another glycogen chain. Each transferred segment must
come from a chain of at least 11 residues, and the new branch point must be at least 4
residues away from other branch points.
 Control of Glycogen Metabolism

The opposing processes of glycogen breakdown and

synthesis are reciprocally regulated by allosteric
interactions and the covalent modification of key
enzymes.

Glycogen metabolism is ultimately under the control

of hormones such as insulin, glucagon, and
epinephrine.
 Glycogen Phosphorylase and Glycogen Synthase Are under Allosteric Control
Both glycogen phosphorylase and glycogen synthase are under allosteric control by effectors that include
ATP, G6P, and AMP.

Muscle glycogen phosphorylase is activated by AMP and inhibited by ATP and G6P. This suggests that when there is
high demand for ATP (low [ATP], low [G6P], and high [AMP]), glycogen phosphorylase is stimulated and glycogen
synthase is inhibited, which favors glycogen breakdown.

Conversely, when [ATP] and [G6P] are high, glycogen synthesis is favored because
glycogen synthase is activated by G6P.

Control by allosteric effectors is superimposed on control by covalent modification.

Control of Glycogen Metabolism
 Enzymes for Phosphate Group Transferring

Kinase is a type of enzyme that transfers phosphate groups from high-energy

donor molecules, such as ATP, to specific substrates, a process referred to as
phosphorylation.

Phosphorylas is an enzyme that catalyzes the addition of a phosphate group

from an inorganic phosphate (phosphate+hydrogen) to an acceptor.

Phosphatase is an enzyme that removes a phosphate group from its substrate

by hydrolysing phosphoric acid monoesters into a phosphate ion and a
molecule with a free hydroxyl group
 Glycogen Phosphorylase Is Activated by Phosphorylation.

Phosphorylase Kinase Is Activated by Phosphorylation

and by Ca2+. Ca2+ concentrations as low as 107 M
activate phosphorylase kinase by inducing a
conformational change in the subunit which is
calmodulin.

The physiological significance of the Ca2+ trigger for this activation is that muscle
contraction is also triggered by a transient increase in the level of cytosolic Ca2+. The rate
of glycogen breakdown is thereby linked to the rate of muscle contraction.
 Glycogen Synthase Is Elaborately Regulated.

The glycogen synthesis and breakdown are linked by PKA and phosphorylase kinase, which, through
phosphorylation, activate glycogen phosphorylase as they inactivate glycogen synthase.
 The adenylate cyclase signaling system

Turning off the Activation of

signal protein kinase A
transduction
Hormonal Control of Glycogen Metabolism
Glycogen metabolism in the liver is largely controlled by the polypeptide hormones insulin and
glucagon acting in opposition. In muscles and various tissues, control is exerted by insulin and
by the adrenal hormones epinephrine and norepinephrine. Glucagon is released from the
pancreas when the concentration of circulating glucose decreases to less than ~5 mM, such as
during exercise or several hours after a meal has been digested. Glucagon is therefore critical
for the liver's function in supplying glucose to tissues that depend primarily on glycolysis for
their energy needs. Muscle cells do not respond to glucagon because they lack the appropriate
receptor.
Epinephrine and norepinephrine, which are often called the fight or flight hormones, are released into
the bloodstream by the adrenal glands in response to stress. There are two types of receptors for these
hormones: the -adrenoreceptor (-adrenergic receptor), which is linked to the adenylate cyclase system,
and the -adrenoreceptor (-adrenergic receptor), whose second messenger causes intracellular [Ca2+] to
increase. Muscle cells, which have the -adrenoreceptor, respond to epinephrine by breaking down
glycogen for glycolysis, thereby generating ATP and helping the muscles cope with the stress that
triggered the epinephrine release. Liver cells respond to epinephrine directly and indirectly because
epinephrine promotes the release of glucagon from the pancreas which activates glycogen phosphorylase
and inactivates glycogen synthase.
Insulin and Epinephrine Are Antagonists. Insulin is released from the pancreas in response to high levels
of circulating glucose (e.g., immediately after a meal). Hormonal stimulation by insulin increases the rate
of glucose transport into the many types of cells that have both insulin receptors and insulin sensitive
glucose transporters called GLUT4 on their surfaces (e.g., muscle and fat cells, but not liver and brain
cells). In addition, [cAMP] decreases, causing glycogen metabolism to shift from glycogen breakdown to
glycogen synthesis by activating phosphoprotein phosphatase-1.

In liver, insulin stimulates glycogen synthesis as a result of the inhibition of glycogen synthase kinase .
This action decreases the phosphorylation of glycogen synthase, thus increasing its activity. In addition, it
is thought that glucose itself, may be a messenger to which glycogen metabolism system responds. An
increase in glucose concentration therefore promotes inactivation of glycogen phosphorylase a through its
conversion to phosphorylase b. The subsequent release of phosphoprotein phosphatase-1 activates
glycogen synthase. Thus when glucose is plentiful, the liver can store the excess as glycogen.
 Comparison of the pathways of gluconeogenesis and glycolysis

Gluconeogenesis

When dietary sources of glucose are not available and when

the liver has exhausted its supply of glycogen, glucose is
synthesized from noncarbohydrate precursors by
gluconeogenesis. In fact, gluconeogenesis provides a
substantial fraction of the glucose produced in fasting
humans, even within a few hours of eating. Gluconeogenesis
occurs in liver and, to a lesser extent, in kidney.

The noncarbohydrate precursors that can be converted to

glucose include the glycolysis products lactate and pyruvate,
citric acid cycle intermediates, and the carbon skeletons of
most amino acids.
 Pyruvate Is Converted to Phosphoenolpyruvate in Two Steps

The first unique reaction of gluconeogenesis

PEP Carboxykinase

Pyruvate carboxylase has a

biotin prosthetic group.

Oxaloacetate is both a precursor for gluconeogenesis and an intermediate of the citric acid
cycle. When the citric acid cycle substrate acetyl-CoAaccumulates, it allosterically
activates pyruvate carboxylase, thereby increasing the amount of oxaloacetate that can
participate in the citric acid cycle. When citric acid cycle activity is low, oxaloacetate
instead enters the gluconeogenic pathway.
 Gluconeogenesis Requires Metabolite Transport between Mitochondria and Cytosol.

The generation of oxaloacetate from

pyruvate or citric acid cycle intermediates
occurs only in the mitochondrion. PEP or
oxaloacetate are then transported across
the mitochondrial membrane.
Oxaloacetate is transported by malate
aspartate shuttle system.

NADH is
transported out of
mitochondria.

mitochondrial
PEPCK
 The second and third unique reactions of gluconeogenesis

Fructose-1,6-bisphosphate is hydrolyzed by
fructose-1,6-bisphosphatase (FBPase). Glucose-
6-phosphate is hydrolyzed by glucose-6-
phosphatase, the same enzyme that converts
glycogen-derived G6P to glucose and which is
present only in liver and kidney.

The net energetic cost of converting two

pyruvate molecules to one glucose molecule by
gluconeogenesis is six ATP equivalents which is
less than the energetic profit (2 ATP) of
converting one glucose molecule to two
pyruvate molecules via glycolysis. The energetic
cost is paid to maintain the independent
regulation of two opposing pathways.
 Phosphofructokinase Is the Major Flux-Controlling Enzyme of Glycolysis in Muscle

In muscle, [ATP] is ~50 times greater than [AMP] and ~10 times greater than [ADP].
Consequently, a change in [ATP] from, for example, 1 to 0.9 mM, a 10% decrease, can result in
a 100% increase in [ADP] (from 0.1 to 0.2 mM) as a result of the adenylate kinase reaction,
and a >400% increase in [AMP] (from 0.02 to ~0.1 mM). Therefore, a metabolic signal
consisting of a decrease in [ATP] too small to relieve PFK inhibition is amplified significantly
by the adenylate kinase reaction, which increases [AMP] by an amount that produces a much
larger increase in PFK activity.

The most potent allosteric effector of PFK is F2,6P, which will be discussed in Section 16-4C.
 Allosteric Effectors Influence Gluconeogenic Flux.

Fructose-2,6-bisphosphate is an extremely potent allosteric activator of

phosphofructokinase (PFK) and an inhibitor of fructose-1,6-bisphosphatase (FBPase).

A bifunctional enzyme functions as both phosphofructokinase-2 (PFK-2) and fructose

bisphosphatase-2 (FBPase-2).
For example, when [glucose] is low, glucagon stimulates the production of cAMP in liver cells.
This activates PKA to phosphorylate the bifunctional enzyme, which inactivates the enzyme's
PFK-2 activity and activates its FBPase-2 activity. The net result is a decrease in [F2,6P], which
shifts the balance between the PFK and FBPase reactions in favor of FBP hydrolysis and hence
increases gluconeogenic flux.
 Regulation by Changing the amounts of enzymes synthesized.

For example, insulin inhibits transcription of the gene for PEPCK, whereas high
concentrations of intracellular cAMP promote the transcription of the genes for
PEPCK, FBPase, and glucose-6-phosphatase, and repress transcription of the genes
for glucokinase, PFK, and the PFK-2/FBPase-2 bifunctional enzyme.