Topic 3:

Glycogen metabolism, metabolic regulation,

citric acid cycle, fatty acid catabolism

Vũ Thanh Ngọc, PhD

Department of Applied Chemistry
School of Biotechnology
HCMC International University
vtngoc@hcmiu.edu.vn
 Metabolism of glycogen in animals
 Excess glucose is converted to polymeric forms for storage:
 Glycogen in vertebrates and many microorganisms
 Starch in plants
 Glycogen is stored in muscle and liver as large particles.
 Contained within the particles are the enzymes that metabolize glycogen,
as well as regulatory enzymes.
 Glycogen metabolism:
 Catabolism: glycogen breakdown
 Anabolism: glycogen synthesis
Glycogen metabolism in animals

1. Glycogen breakdown (catabolism)

2. Glycogen synthesis (anabolism)

Glycogen structure
 Glycogen breakdown

In skeletal muscle and liver, the glucose units of the outer branches of
glycogen enter the glycolytic pathway through the action of 3 enzymes:
 Glycogen phosphorylase
 Glycogen debranching enzyme
 Phosphoglucomutase
 Glycogen breakdown
 Glycogen phosphorylase (1)
 Glycogen debranching enzyme (2)
 Phosphoglucomutase (3)

(1) Glycolysis
+
(2) (3)
Glycogen G-1-P G-6-P

Glucose-6-phosphatase
(ONLY in liver, kidney) Glucose
Glycogen breakdown
Glycogen breakdown
Glycogen breakdown
 Glycogen breakdown
 Glycogen phosphorylase (1)
 Glycogen debranching enzyme (2)
 Phosphoglucomutase (3)

(1) Glycolysis (cytosol)

+
(2) (3)
Glycogen G-1-P G-6-P

Glucose-6-phosphatase
(ONLY in liver, kidney) Glucose

Does G-6-P to glucose reaction take place in cytosol?

 Glycogen breakdown
Hydrolysis of glucose 6-phosphate by glucose 6-phosphatase of the ER

Genetic defects in either glucose 6-phosphatase or T1 lead to serious

derangement of glycogen metabolism, resulting in type Ia glycogen storage
disease
 Glycogen synthesis
Glycogen synthesis takes place in virtually all animal tissues but is
especially prominent in the liver and skeletal muscles.

phosphoglucomutase UDP-glucose pyrophosphorylase

G-6-P G-1-P + UTP UDP-Glucose + PPi

glycogen synthase

transfer glucose residue from UDP-glucose to

a nonreducing end of a branched glycogen
 Glycogen synthesis
Branch synthesis in glycogen by glycogen-branching enzyme (glycosyl-
(46)-transferase)
 Glycogen synthesis

Glycogen synthase cannot initiate a new glycogen chain de novo.

 Glycogenin to make a primer
Glycogen synthesis

Glycogenin and the structure

of the glycogen particle.
Glycogen synthesis

Glycogenin and the structure

of the glycogen particle.
 Metabolic regulation

1. Regulation of metabolic pathways

2. Coordinated regulation of glycolysis and gluconeogenesis

3. Coordinated regulation of glycogen synthesis and breakdown

Regulation of metabolic pathways

 The role of metabolic regulation

 Bioenergetic view of metabolic regulation

 Mechanistic regulation of enzyme activity

Regulation of metabolic pathways
 The role of metabolic regulation
 Maintain homeostasis at the molecular, cellular, and organismal level

 Maintain a dynamic steady state

 When steady state is disturb

 Trigger regulatory mechanisms

 Make adjustments

 Return the organism to a new steady state

 Achieve homeostasis
 Regulatory mechanisms evolved under
strong selective pressures

 Maintain homeostasis

 Maximize the efficiency of fuel utilization by preventing the simultaneous

operation of pathways in opposite directions (such as glycolysis and
gluconeogenesis)
 Regulatory mechanisms evolved under
strong selective pressures
 Partition metabolites appropriately between alternative pathways (such as
glycolysis and the pentose phosphate pathway)

 Draw on the fuel best suited for the immediate needs of the organism
(glucose, fatty acids, glycogen, or amino acids)

 Shut down biosynthetic pathways when their products accumulate

Bioenergetic view of metabolic regulation

The most important purpose of metabolic regulation is to maintain a

constant supply and concentration of ATP

ATP  ADP or AMP

 regulatory mechanisms responding to change in [ATP]/[ADP] ratio

Bioenergetic view of metabolic regulation
ATP  ADP or AMP

[AMP] is a much more sensitive indicator of a cell’s energetic state

than is [ATP]

 Many regulatory processes hinge on changes in [AMP]

 Bioenergetic view of metabolic regulation
Maintain a dynamic steady state

 to hold some cellular parameter (Ex: concentration of a metabolite) at a

steady level over time, even as the flow (rate) of metabolites through the
pathway changes.

 Regulate the rate of metabolite flow (flux) so that the concentration

of substrate is constant
Bioenergetic view of metabolic regulation
Flux through a biochemical pathway depends on the activities of the
enzymes that catalyze each reaction in that pathway

 Regulate enzyme activity

(1) What changes in the reaction do regulatory enzymes respond to?

? (2) Which reactions (enzymes) in the pathway are under regulation?

(3) How to regulate enzyme activity?

 Bioenergetic view of metabolic regulation

(1) What changes in the reaction do regulatory enzymes respond to?

 Bioenergetic view of metabolic regulation

 Mass action ratio, Q: the ratio of products to substrates

 if [ADP]/[ATP] is to be maintained, [G6P]/[Glc] becomes important.
 Q is dictated by metabolite concentration.
 Regulatory enzymes respond to changes in metabolite concentration
 Bioenergetic view of metabolic regulation

(2) Which reactions (enzymes) in the pathway are under regulation?

Bioenergetic view of metabolic regulation

Discussion:
Near-equilibrium:
 How is actual free-energy change?
 How is Q relative to K’eq?
 Bioenergetic view of metabolic regulation
Reactions are near equilibrium, nothing to be regulated about.
The metabolic regulation is to keep reactions with very large
differential between Q and K’eq or large free energy change to
stay away from equilibrium.

Discussion:
Should the reaction ATP to ADP and Pi be allowed to approach
equilibrium in cells? Why?
Bioenergetic view of metabolic regulation
Near-equilibrium and non-equilibrium steps in a metabolic pathway

 Enzyme activity in the non-equilibrium steps is under tight regulation

Non-equilibrium: free-energy change, differential between Q and K’eq?

Bioenergetic view of metabolic regulation

(1) What changes in the reaction do regulatory enzymes respond to?

(2) Which reactions (enzymes) in the pathway are under regulation?

(3) How to regulate enzyme activity?

 Mechanistic regulation of enzyme activity

Enzyme activity can be regulated by:

 Changing the rate of its synthesis or degradation,

 Allosteric or covalent alteration of existing enzyme molecules,

 Separating the enzyme from its substrate in subcellular compartments.

Mechanistic regulation of enzyme activity
Different types of covalent modification

 Adenylylation

 Methylation

 Attachment of lipids

 Phosphorylation: most common

Coordinated regulation of glycolysis and gluconeogenesis

 Regulation of glycolysis: 3 regulatory enzymes

 Regulation of glyconeogenesis: 2 control points

 Reciprocal allosteric control of the 2 pathways by fructose 2,6-bisphosphate

 Glycolysis vs gluconeogenesis
GLYCOLYSIS
5
HK PHI PFK1 A
1 2 3 4 TPI
G G6P F6P F16BP G3P + DHAP
6 G3PD
G6P FBPase1
13PG
GLUCONEOGENESIS 7 PGK
3PG
8 PGM
7 reversible reactions 2PG
3 irreversible bypass reactions 9 E
PEP
Oxaloacetate 10 PK
Pyruvate
 Regulation of glycolytic enzymes

3 glycolytic enzymes are subjected to allosteric regulation:

 Hexokinase (HK)

 Phosphofructokinase-1 (PFK-1)

 Pyruvate kinase (PK)

 Regulation of glycolytic enzymes: (1) Hexokinase

Hexokinase (HK), regulatory enzyme, has 4 isozymes:

 Myocytes: HK II

 Muscle: HK I-III - allosterically inhibited by glucose 6-phosphate

 Liver: HK IV
 Regulation of glycolytic enzymes: Hexokinase

HK IV (liver) is different from HK I-III:

 Half-saturated at higher glucose concentration

 Inhibited by the reversible binding of a regulatory protein specific to liver

(nucleus sequestration)

 NOT inhibited by glucose 6-phosphate

 Regulation of glycolytic enzymes: Hexokinase

Regulation of HK IV by sequestration in the nucleus.

Regulation of glycolytic enzymes: (2) Phosphofructokinase-1

 PFK-1 is allosterically inhibited by ATP and citrate.

 PFK-1 is allosterically activated by fructose 2,6-bisphosphate.

 Regulation of glycolytic enzymes: (3) Pyruvate kinase
 PK is allosterically inhibited by ATP
 The liver isozyme is inhibited by cAMP-dependent phosphorylation
 Regulation of glyconeogenesis: 2 control points

2 control points of glyconeogenesis:

 Fate of pyruvate:

citric acid cycle or gluconeogenesis

 Activity of FBPase-1
Regulation of glyconeogenesis: fate of pyruvate
Regulation of glyconeogenesis: activity of FBPase-1
Reciprocal allosteric control of glycolysis and gluconeogenesis
by fructose 2,6-bisphosphate (F26BP)
F26BP activates PFK-1 and stimulates glycolysis in liver and, at the same
time, inhibits FBPase-1, thereby slowing gluconeogenesis
Reciprocal allosteric control of glycolysis and gluconeogenesis
by fructose 2,6-bisphosphate

Cellular [F26BP] depends on its synthesis by phosphofructokinase-2 (PFK-2)

and breakdown by fructose 2,6-bisphosphatase (FBPase-2).
 Reciprocal allosteric control of glycolysis and gluconeogenesis
by fructose 2,6-bisphosphate
PFK-2 and FBPase-2 are 2
parts of the same protein
and both are regulated, in
a reciprocal fashion, by
insulin and glucagon.

Phosphorylation of PFK-
2/FBPase-2 enhances its
FBPase-2 activity and
inhibits its PFK-2 activity
(vice versa)
Coordinated regulation of glycogen synthesis and breakdown:
glycogen phosphorylase activity

Glycogen phosphorylase
Glycogen G-1-P

Glycogen phosphorylase exists in 2 interconvertible forms:

 Glycogen phosphorylase a (active)
 Glycogen phosphorylase b (less active)
 Coordinated regulation of glycogen synthesis and breakdown:
glycogen phosphorylase activity

Glycogen phosphorylase is regulated allosterically and hormonally

 Allosterically by Ca2+, AMP and glucose
 Hormonally by covalent modification
(phosphorylation/ dephosphorylation)
 Coordinated regulation of glycogen synthesis and breakdown:
glycogen phosphorylase activity

Regulation of muscle glycogen phosphorylase

by covalent modification
Coordinated regulation of glycogen synthesis and breakdown:
glycogen phosphorylase activity

Cascade mechanism of epinephrine

and glucagon action
Coordinated regulation of glycogen synthesis and breakdown:
glycogen phosphorylase activity

Glycogen phosphorylase of liver as a glucose sensor

Coordinated regulation of glycogen synthesis and breakdown:
glycogen synthase activity

Glycogen synthase
UDP-Glucose Glycogen elongation

Glycogen synthase exists in phosphorylated/unphosphorylated form:

 Glycogen synthase a (active): unphosphorylated
 Glycogen synthase b (inactive): phosphorylated (allosterically
activated by G6P)
 Coordinated regulation of glycogen synthesis and breakdown:
glycogen synthase activity

Glycogen synthase
UDP-Glucose Glycogen elongation

 Glycogen synthase is phosphorylated by many protein kinases.

 The most important regulatory kinase is glycogen synthase kinase 3 (GSK3)
Coordinated regulation of glycogen synthesis and breakdown:
glycogen synthase activity

Effects of GSK3 on glycogen synthase activity

Coordinated regulation of glycogen synthesis and breakdown:
glycogen synthase activity
GSK3 mediates the actions of insulin
on glycogen synthase activity
Coordinated regulation of glycogen synthesis and breakdown:
PP1 is central to glycogen metabolism

PP1 can remove phosphoryl groups from all 3 enzymes phosphorylated

in response to glucagon (liver) and epinephrine (liver and muscle):
 Phosphorylase kinase
 Glycogen phosphorylase
 Glycogen synthase
 Coordinated regulation of glycogen synthesis and breakdown:
Allosteric & hormonal signals coordinate carbohydrate metabolism

Regulation of carbohydrate metabolism in the hepatocyte

 Coordinated regulation of glycogen synthesis and breakdown:
Allosteric & hormonal signals coordinate carbohydrate metabolism

Difference in the regulation of carbohydrate metabolism in liver and muscle

Coordinated regulation of glycogen synthesis and breakdown:
Insulin changes the expression of many genes
involved in carbohydrate and fat metabolism
SUMMARY: GLYCOGEN METABOLISM

Glycogen metabolism in animals

 Glycogen synthesis

 Glycogen breakdown
 SUMMARY: METABOLIC REGULATION
Regulation of metabolic pathways
 The role of metabolic regulation
 Bioenergetic view of metabolic regulation
 Mechanistic regulation of enzyme activity

Coordinated regulation of glycolysis and gluconeogenesis

 Regulation of glycolysis: 3 regulatory enzymes
 Regulation of glyconeogenesis: 2 control points
 Reciprocal allosteric control of the 2 pathways by F26BP
 SUMMARY: METABOLIC REGULATION

Coordinated regulation of glycogen synthesis and

breakdown:
 Regulation of glycogen phosphorylase activity

 Regulation of glycogen synthase activity

 Allosteric & hormonal signals coordinate carbohydrate metabolism

 Insulin changes the expression of many genes involved in carbohydrate

and fat metabolism