Biochemistry 2

Storage
Mechanisms and
Control in
Carbohydrate
Metabolism
 Carbohydrates

• The “quick energy” that is needed by our

body.
• When we eat carbs, our body breaks them
down into simple sugars.
• The body can mobilize carbohydrates more
easily than fat.
 Glycogen

• Rapid energy source.

• The stored form of glucose that is made of
connected glucose molecules.
• Its structure is optimized for its ability to
store and deliver energy quickly.
• The average chain length of its branches
are 13 residues.
How is Glycogen Produced and Degraded?
 Glycogenolysis:
The Breakdown of Glycogen
Glycogenolysis (Breakdown of Glycogen)

First Reaction: Phosphorolysis reaction catalyzed by

Glycogen Phosphorylase
Glycogenolysis (Breakdown of Glycogen)

Second Reaction: Isomerization; catalyzed by

Phosphoglucomutase
Glycogenolysis (Breakdown of Glycogen)

• Glycogen phosphorylase cleaves the

α (1 4) linkages.
• No ATP is hydrolyzed to ADP in this
reaction.
• An alternative mode of entry to
glycolytic pathway which saves one ATP
for each molecule of glucose. 1st Step of Glycolysis

• Net gain of 3 ATP per glucose.

Glycogenolysis (Breakdown of Glycogen)

Third Reaction: Debranching; catalyzed by Debranching

Enzymes

Debranching Enzymes:

• Transferase – moves 3 residues (limit branch) to the

free end of the linear linkage

• Glucosidase – hydrolyzes the α (1 6) remaining

glucose residue that is linked to the
branch point
Glycogenolysis: Third Reaction (Debranching)

Transferase
Glycogenolysis: Third Reaction (Debranching)

Glucosidase
 Glycogenesis:
The Formation of Glycogen
from Glucose
Glycogenesis (Formation of Glycogen from Glucose)

• Glycogenesis requires energy which is provided by the

hydrolysis of uridine triphosphate (UTP).
• In the first stage of glycogen synthesis, glucose-1-phosphate
reacts with UTP to produce uridine diphosphate glucose
(UDP-glucose or UDPG) and pyrophosphate.
Glycogenesis (Formation of Glycogen from Glucose)

UDP-glucose phosphorylase

+
∆G ≈ 0

Pyrophosphate
Glycogenesis (Formation of Glycogen from Glucose)

Pyrophosphatase

∆G = -30.5 kJ/mol
Glycogenesis (Formation of Glycogen from Glucose)
Glycogenesis (Formation of Glycogen from Glucose)

• The supply of UTP is replenished by an exchange

reaction with ATP, which is catalyzed by
nucleoside phosphate kinase
Glycogenesis (Formation of Glycogen from Glucose)

• The addition of UDPG to a growing chain of glycogen is the

next step in glycogen synthesis.

• The hydroxyl group of a specific tyrosine of the protein

glycogenin acts as a primer which initiates the glycogen
synthesis.

• Glycogenin acts as a catalyst for addition of glucoses until

there are about eight of them linked together.
Glycogenesis (Formation of Glycogen from Glucose)

At least 8 glucose molecules are linked before

glycogen synthase takes over.
Glycogenesis (Formation of Glycogen from Glucose)

• Glycogen synthase catalyzes the addition of α (1 4)

glucose linkages to the existing chain.

• Glycogenesis also requires the formation of α (1

6) in
which a branching enzyme accomplishes this task.
 Glycogenesis (Formation of Glycogen from Glucose)

• The branching enzyme

catalyzes the transfer of 7
residues from the end of
the growing chain to a
branch point in which it
also catalyzes the
formation of the required
α (1 6) glycosidic linkage.
 Glycogenesis (Formation of Glycogen from Glucose)

• The transferred 7 residues

must come from a chain
of at least 11 residues
long.
 Glycogenesis (Formation of Glycogen from Glucose)

• The new branch point

must be at least 4
residues away from the
nearest existing branch
point.
Control of Glycogen
Metabolism
How Does Gluconeogenesis Produce
Glucose from Pyruvate?
 Glucose

G-6-P Pyruvate
CO2
F-6-P

F-1,6-BP

DHAP GA-3-P

1,3-BPG

Cytosol 3-PG

2-PG

PEP

Mitochondria
Pyruvate
Gluconeogenesis: Production of Glucose from Pyruvate

First Reaction: Catalyzed by Pyruvate carboxylase

First Reaction: Pyruvate + CO2 Oxaloacetate

• Requires energy which is provided by the hydrolysis of ATP.

• Pyruvate carboxylase is an allosteric enzyme found in
mitochondria.
• Acetyl-CoA is an allosteric effector that activates the enzyme.
• Magnesium ion and biotin are also required for an effective
catalysis.
• Biotin is a carrier of carbon dioxide.
• The carboxyl group of biotin forms an
amide bond with the ϵ-amino group of a
specific lysine chain of pyruvate
carboxylase.
 Pyruvate

Carboxylate
dissolved in H2O

Pyruvate
Carboxylase

Oxaloacetate

• CO2 is attached to biotin, in which biotin is covalently bonded to

pyruvate carboxylase.
• After CO2 is attached to biotin, it then shifts to pyruvate to form
oxaloacetate.
• ATP is required for this reaction.
Mitochondria

• The oxaloacetate formed in

mitochondria can have two fates:
1. Leave the mitochondria via a
specific transporter to continue
gluconeogenesis in the cytosol.
2. It can be turned to malate via
mitochondrial malate
dehydrogenase.
Gluconeogenesis: Production of Glucose from Pyruvate

Second Reaction: Catalyzed by Phosphoenolpyruvate carboxykinase (PEPCK)

Second Reaction: Oxaloacetate Phosphoenolpyruvate

• Phosphoenolpyruvate carboxykinase (PEPCK) is found in the

mitochondria and cytosol.
• The reaction involves hydrolysis of GTP.
• The successive carboxylation and decarboxylation reactions are
both close to equilibrium.
 Glucose

G-6-P

F-6-P

Mitochondria
F-1,6-BP

DHAP GA-3-P

1,3-BPG

3-PG
Cytosol
2-PG

PEP

Pyruvate
Role of Sugar Phosphates in

Gluconeogenesis
Gluconeogenesis: Production of Glucose from Pyruvate

First Reaction: Catalyzed by Fructose-1,6-bisphosphatase

First Reaction: Fructose-1,6-bisphosphate Fructose-6-phosphate

• Fructose-1,6-bisphosphatase is an allosteric enzyme strongly

inhibited by AMP but stimulated by ATP.
• This enzyme is also inhibited by fructose-2,6-bisphosphate (a
potent activator of phosphofructokinase).
Gluconeogenesis: Production of Glucose from Pyruvate

Second Reaction: Catalyzed by Glucose-6-phosphatase

Second Reaction: Glucose-6-phosphate Glucose

• The hydrolysis of glucose-6-phosphate to glucose occurs in the

endoplasmic reticulum.
 Glucose

G-6-P
Mitochondria
F-6-P

F-1,6-BP

DHAP GA-3-P

1,3-BPG

3-PG
Cytosol

Endoplasmic Reticulum 2-PG

PEP

Pyruvate
How Is Carbohydrate Metabolism
Controlled?
Control of Phosphofructokinase and Fructose-1,6- biphosphatase
Glycolysis–Gluconeogenesis: reciprocal regulated
Regulation of glycolysis To bloodstream Regulation of
gluconeogenesis

GLUCOSE

- Glucose-6-phoshate Hexokinase Glucose-6-phoshatese

GLUCOSE-6-PHOSPHATE

FRUCTOSE-6-PHOSPHATE
+ Fructose-2,6-biphosphate
- F-2,6-BP
+ AMP Phosphofructokinase Fructose-1,6-biphosphatase
- ATP - AMP
- Citrate FRUCTOSE-1,6-BIPHOSPHATE

PHOSPHENOLPYRUVATE
Phosphoenolpyruvate
+ Fructose-1,6-biphosphate carboxykinase
- Acetyl-CoA Pyruvate Kinase
OXALOACETATE
- ATP
Pyruvate carboxylase + Acetyl-CoA

PRUVATE
 Control of Phosphofructokinase and Fructose-1,6-
biphosphate
• Involves fructose-2,6-bisphosphate (F2,6P).
 An allosteric activator of phosphofructokinase (PFK).
 An inhibitor of fructose bisphosphate phosphatase. (FBPase)
• Concentration of fructose-2,6-bisphosphate :
 high stimulates glycolysis.
 low stimulates gluconeogenesis.
 Control of Phosphofructokinase and Fructose-1,6-
biphosphate

• The concentration of F2,6P in a cell depends on the balance between its synthesis, catalyzed by
phosphofructokinase-2 (PFK-2), and its breakdown, catalyzed by fructose-bisphosphatase-2 (FBPase-
2).
 Control of Phosphofructokinase and Fructose-1,6-
biphosphate

• The inhibitor works by itself, but its effect is

greatly increased by thepresence of the
allosteric inhibitor AMP.
Mechanism of Metabolic Control
Type of Control Mode of Operation Examples

Allosteric Effectors (substrates, products, or coenzymes) of a ATCase (Section 7.2);

pathway phosphofructokinase (Section 17.2)
inhibit or activate an enzyme. (Responds rapidly to
external stimuli.)

Covalent modification Inhibition or activation of enzyme depends on Sodium–potassium pump (Section 8.6);
formation glycogen phosphorylase, glycogen
or breaking of a bond, frequently by phosphorylation synthase (Section 18.1)
odephosphorylation. (Responds rapidly to external
stimuli.)

Substrate cycles Two opposing reactions, such as formation and Glycolysis (Chapter 17) and
breakdown of a given substance, are catalyzed by gluconeogenesis (Section 18.2)
different enzymes, which can be activated or
inhibited separately. (Responds
rapidly to external stimuli.)

Genetic control The amount of enzyme present is increased by Induction of b-galactosidase (Section
protein synthesis. (Longer-term control than the 11.2)
other mechanisms listed here.)
 Glycogen Metabolism in Body Organ

• Using combinations of these control mechanisms, an organism can set up a

division of labor among tissues and organs to maintain control of glucose
metabolism.

• In the same cell (of whatever type), these two metabolic pathways (glycolysis
and gluconeogenesis) are not highly active simultaneously.
Glycogen Metabolism in Body Organ
Glycogen Metabolism in Body Organ
 Control of Pyruvate Kinase
• The final step of glycolysis is also a major control point in glucose metabolism;
1) Pyruvate kinase (PK) is allosterically affected (Inhibitors: ATP and alanine)
2) ) Alanine is the amino version of pyruvate (by transaminase).
 A high level of alanine
 A high level of pyruvate
 Making pyruvate: shut down
3) Fructose-1,6-bisphosphate allosterically activates PK so that the incoming products of
the first reactions of glycolysis can be processed.
Control of Pyruvate Kinase

4.) Pyruvate kinase is also found as isozymes (subunits: M, L, A).

 M subunit, in muscle; L, in liver; A, in other tissues.
 Liver isozymes subject to covalent modification and allosteric
control.
 Low levels of blood sugar trigger the release of
glucagons production of a protein kinase.
Control of Pyruvate Kinase
Why Is Glucose Sometimes Diverted through the
Pentose Phosphate Pathway
Pentose phosphate pathway

• five-carbon sugars (including ribose) are

produced from glucose.
• Production of nicotinamide adenine
dinucleotide phosphate (NADPH)
 NADH is produced in the oxidative
reactions
 NADPH is a reducing agent in
biosynthesis
Pentose phosphate pathway: oxidative
• Glucose-6-phosphate is oxidized to 6-phosphogluconate.

\]’’]NADPH is produced by the reaction.

• Oxidative decarboxylation, and NADPH is produced once again.
 6-Phosphogluconate  loses its carboxyl group (released as
CO2), production of ribulose-5-phosphate.
 Enzyme: 6-phosphogluconate dehydrogenase.
Pentose phosphate pathway: oxidative
Pentose phosphate pathway: nonoxidative reactions

• In the remaining steps of the pentose phosphate pathway, several

reactions involve transfer of 2- and 3-carbon units.
• Two different reactions in which ribulose-5-phosphate isomerizes.
 Phosphopentose-3-epimerase
 Phosphopentose isomerase
Pentose phosphate pathway: nonoxidative reactions

• Group-transfer reactions require the two five-carbon sugars produced by

the isomerization of ribulose-5-phosphate.
• 2 molecules of xylulose-5-phosphate and 1 molecule of ribose-5-
phosphate rearrange to give 2molecules of F6P and 1 molecule of
glyceraldehyde-3-phosphate.
 3 molecules of pentose (C5) give 2 molecules of hexose (C6)
and 1 molecule of a triose (C3).
Pentose phosphate pathway: nonoxidative reactions
• Two enzymes that are responsible for the reshuffling of the carbon
atoms of sugar;
 Transketolase
 Transaldolase
Pentose phosphate pathway: regulation

• The reactions catalyzed by transketolase and transaldolase are reversible.

• If the organism needs more NADPH than ribose-5-phosphate
• .If the organism has a greater need for ribose-5-phosphate than for NADPH
Pentose phosphate pathway: regulation
 2022 March
27

Thank you
for
listening!
For
Presentation