CARBOHYDRATE METABOLISM

CATABOLISM

EDITED BY
Liniyanti D.Oswari,MD.,MNS,MSc.
For Block 7
2019

Medical student, Sriwijaya University

 Metabolism
-Metabolism can be organized into pathways: these pathways
may overlap, and branch, and even consist of circular paths

- A pathway consists of consecutive enzymatic reactions: the

reactants, intermediates and products are called metabolites

- The flux through each pathway is regulated by a

sophisticated set of metabolic regulatory mechanisms:
feedback inhibition, allosteric interactions, covalent
modification of proteins/enzymes, etc.

- Over a 40 year time span a human being consumes tons of

nutrients and imbibes 20,000 liters of water, yet remains more
or less at steady state
-Metabolic pathways are irreversible at certain key steps
where a large negative free energy change occurs
-There are catabolic pathways (e.g.glycolysis) and anabolic
pathways (e.g. gluconeogenesis);these are necessarily different
at key steps, but may share some steps
-In eukaryotic cells the various metabolic pathways are
compartmentalized:cytosol, mitochondria, peroxisomes,
chloroplasts, etc.
-The compartments are defined by membranes; special
transport mechanisms mustexist to regulate the flow of
reactants and products in and out of a compartment
-In some cases the flow is determined by a concentration
gradient (diffusion), but stillrestricted by a specific transporter;
in other cases integral membrane proteins can act as pumps to
pump metabolites and ions against a concentration gradient;
this requires an input of energy
Major Pathways of CHO Metabolism
CHO metabolism in mammalian cells can be classified
into:
1. Glycolysis: Oxidation of glucose to pyruvate (aerobic
state) or lactate (anaerobic state)
2. Krebs cycle: After oxidation of pyruvate to acetyl
CoA, acetyl CoA enters the Krebs cycle for the aim of
production of ATP.
3. Hexose monophosphate shunt: Enables cells to
produce ribose-5-phosphate and NADPH.
4. Glycogenesis: Synthesis of glycogen from glucose,
when glucose levels are high
5. Glycogenolysis: Degradation of glycogen to glucose
when glucose in short supply. Augus 27
4
6. Gluconeogenesis: Formation t 2019of glucose from
Dr. Mohamed Z Gad mohamed.gad@guc.edu.eg
Glucose is the major fuel of most organisms. The
major pathways of CHO metabolism either begin
or end with glucose.
Carbohydrate Metabolism
 Glycolysis
 2.3. Biphospoglycerate (2.3.BPG)
 Glycogenesis
 Glycogenolysis
 HMP shunt
 Gluconeogenesis
 REGULATION OF METABOLISM BY
HORMONES
Carbohydrate Metabolism Overview
Glycogen

pentose GLUCOSE other

sugars

pyruvate

acetyl CoA EtOH

lactate
TCA cycle ATP
 Glucose Utilization

Energy
Adipose Stores Glycogen

Glucose
Pentose Glycolysis
Phosphate
Pathway

Ribose-5-phosphate Pyruvate
GLYCOLYSIS
 Glucose can also be available from food
intake.
 Glucose is also stored as glycogen
(glycogenesis).
 After gluconeogenesis, glucose is
converted from glycogen in liver or muscle
for glycolysis.
 Glycolysis is the break down of a 6 C
glucose sugar to two 3C pyruvate.
Central role of liver in metabolism

 Glucose entering the hepatocyte is

phosphorylated by glucokinase to glucose-6-
phosphate (G-6-P).
 Other monosaccharides are also made to G-6-P
via gluconeogenesis, then glucose can be stored
as glycogen.
 When we need energy, glycolysis converts G-6-P
to pyruvate and acetyl coA to enter Citric acid
cycle to produce ATP energy via oxidative
phosporylation (aerobic metabolism).
Glycolysis:
break down of glucose in cytoplasm
UDP-glucose
Glucose-1-
phosphate
Glycogen

Lactate
Dehydrogenase
Glucose-6- Hexokinase
phosphate
Glucose

Lactate
ADP ATP
Fructose-6-phosphate

Pyruvate
ADP
ATP
Fructose-1, 6-biphosphate

Glyceraldehyde-3- Dihydroxyacetone Glycerol ADP ATP

phosphate phosphate (DHAP)
NAD + Pi
ADP ATP H2O Phospho
NADH + H+
Glyceraldehyde-1, Glycerate-3- Glycerate-2- -enol-
3-bisphosphate phosphate phosphate pyruvate
ADP ATP H2O
Glycolysis: Phase 1 and 2
 Phase 1: Sugar activation
 Two ATP molecules activate glucose into
fructose-1,6-diphosphate
The 1 and 6 indicate which carbon atom
to which they are attached.
 Phase 2: Sugar cleavage (splitting)
 Fructose-1,6-bisphosphate(6 C’s) is split
into two 3-carbon compounds:
Glyceraldehyde 3-phosphate (GAP)
Glycolysis: Phase 3
 Phase 3: Oxidation and ATP formation
 The3-carbon sugars are oxidized (reducing
NAD+); i.e., 2 H’s + NAD NADH2
 Inorganicphosphate groups (Pi) are attached
to each oxidized fragment
 The terminal phosphates are cleaved and
captured by ADP to form four ATP molecules
 The final products are:
Two pyruvic acid molecules
Two NADH + H+ molecules (reduced NAD+)
A net gain of two ATP molecules
Glycolysis has two stages.
A. An energy investment phase.
Reactions, 1-5. Glucose to two
glyceraldehyde -3-phosphate
molecules. 2 ATPs are invested.
B. An energy payoff phase.
Reactions 6-10. two glyceraldehyde
3-phosphate molecules to two
pyruvate plus four ATP molecules.
-- A net of two ATP molecules overall
plus 2 NADH(10 ATP–2 ATP= 8 ATP).
 Glycolysis: spend a little get a lot
 For each molecule of glucose:
 2 ATP molecules are spent in the
activation/preparatory phase, but 4 ATP
molecules are produced in the payoff
phase
 The final product, pyruvate is then further
metabolised
 In the next slides we will concentrate on
the cleavage (lysis, that gives this
pathway its name) of Fructose 1,6-
Biphosphate into two smaller molecules:
Glyceraldehyde and Dihydroxyacetone
(the enzyme doing this is the fructose 1,6-
biphosphate aldolase)

15
GLYCOLYSIS Glucose
ATP
hexokinase ADP
Glucose 6-phosphate
phosphogluco-
isomerase
Fructose 6-phosphate
ATP
phosphofructokinase ADP
Fructose 1.6-bisphosphate
aldolase

triose phosphate isomerase

Dihydroxyacetone Glyceraldehyde
phosphate 3-phosphate
 Glyceraldehyde 3-phosphate
glyceraldehyde NAD+ + Pi
3-phosphate NADH + H+
dehydrogenase
1,3-Bisphosphoglycerate
ADP
phosphoglycerate kinase ATP
3-Phosphoglycerate
phosphoglyceromutase
2-Phosphoglycerate
enolase H2O
Phosphoenolpyruvate
ADP
pyruvate kinase ATP
Pyruvate
Three irreversible kinase reactions
primarily drive glycolysis forward.

 hexokinase or glucokinase
 phosphofructokinase
 pyruvate kinase

These enzymes will be shown to be

regulate glycolysis as well.
 Hexokinase Vs Glucokinase
Hexokinase Glucokinase
Site Most tissues Hepatocytes
Islet cells (pancreas)
Kinetics Low Km High Km
Low Vmax High Vmax
Regulation G-6-phosphate F-6-phosphate
Insulin: Induction
Function Low glucose conc. High glucose conc.
Glucose sensor
-- REGULATION OF GLYCOLYSIS

1.HEXOKINASE and GLUCOKINASE

HEXOKINASE
 Commiting step in glycolysis: phosphorylation
of glucose.
 Inhibited by its product, glucose6-phosphate,
as a response to slowing of glycolysis .
 found in all cells of every organism low
specificity for monosaccharides
(simple sugars) i.e., other monosaccharides
can be phosphorylated by hexokinase.
 relatively high affinity for glucose,
KM = 0.1 mM
GLUCOKINASE

liver enzyme with high KM (10 mM)for

glucose so most effective when
glucose levels are very high
 not inhibited by glucose 6-phosphate
sensitive to high glucose in
circulation from recent meal
 so it decreases high level of glucose
in blood by taking glucose into liver
Hexokinase IV Regulation in Liver
Glycogen
2. PHOSPHOFRUCTOKINASE
 rate limiting for glycolysis
 an allosteric multimeric regulatory
enzyme.
 Measures adequacy of energy levels.

 Inhibitors: ATP and citrate

high energy
 Activators: ADP, AMP, and
fructose 2,6 bisphosphate
low energy
 ATP inhibits phosphofructose
activity by decreasing fructose
6-phosphate bindingAMP and ADP
reverse ATP inhibition
 Fructose 2,6 bisphosphate is a very
important regulator, controlling the
relative flux of carbon through
glycolysis versus gluconeogenesis.
- It also couples these pathways to
hormonal regulation.
3. PYRUVATE KINASE
PEP + ADP Pyruvate + ATP
 An allosteric tetramer
-inhibitor: ATP & acetyl CoA & fatty
acids (alternative fuels for TCA cycle)
- activator: fructose 1,6-bisphosphate
- (“feed-forward”)
 Phosphorylation (inactive form) and
dephosphorylation (active form)
under hormone control.
Also highly regulated at the level of gene
expression(“carbohydrate loading”)
Glycolysis:
Embden-Myerhof
Pathway

 Oxidation of
glucose
 Products:
 2 Pyruvate
 2 ATP
 2 NADH
 Cytosolic
Aerobic Vs Anaerobic Glycolysis
Aerobic Glycolysis:
Total Vs Net ATP Production
Summary of Energy Relationships for
Glycolysis aerobic
Input = 2 ATP
1. glucose + ATP  glucose-6-P
2. fructose-6-P + ATP  fructose
1,6bisphosphate
Output = 4 ATP + 2 NADH
1. 2 glyceraldehyde-3-P + 2 Pi + 2 NAD+
2 (1,3 bisphosphoglycerate) + 2 NADH
2. 2 (1,3 bisphosphoglycerate) + 2 ADP
2 (3-P-glycerate) + 2 ATP
3. 2 PEP + 2 ADP  2 pyruvate + 2 ATP
 Energy Yield From Glycolysis

Glucose 6 CO2 = -2840 kJ/mole

2 ATPs produced = 2 x 30.5 =

61 kJ/mole glucose

Energy yield = 61/2840 = 2%

recovered as ATP
- subsequent oxidation of pyruvate and
NADH can recover more of the free
Carbohydrate Metabolism
 Primarily glucose
 Fructose and galactose enter the pathways at various
points
 All cells can utilize glucose for energy production
 Glucose uptake from blood to cells usually mediated by
insulin and transporters
 Liver is central site for carbohydrate metabolism
 Glucose uptake independent of insulin
 The only exporter of glucose
Blood Glucose Homeostasis
 Several cell types prefer glucose as energy
source (ex., CNS)
 70-110 mg/dl is normal range of
fasting blood glucose
Uses of glucose:
 Energy source for cells
 Muscle glycogen
 Fat synthesis if in excess of needs
Blood glucose levels at
various stages of fasting:
Stage of fasting Glucose (mg/dL) Glucose (mM/L)

Normal level 80-100 4,4-5,6

Fasting (12 h) 80 4,4

Starvation (3 d) 70 3,9

Starvation (5-6 wk) 65 3,6

Sources of blood glucose in fed,
fasting, and starved states:
 Fates of Glucose
 Fed state Synthesis and
 Storage as glycogen breakdown occur
 Liver at all times
 Skeletal muscle regardless of
 Storage as lipids
state...
 Adipose tissue

 Fasted state
The relative rates
 Metabolized for energy
of synthesis and
breakdown change
 New glucose synthesized
 High Blood Glucose

Pancreas

Muscle Insulin Glycogen

Glucose Glucose
absorbed Adipose absorbed
Cells
Glucose absorbed
immediately after eating a meal…
Glucose Metabolism
 Four major metabolic pathways:
 Immediate source of energy
 Pentophosphate pathway
 Glycogen synthesis in liver/muscle
 Precursor for triacylglycerol synthesis
 Energy status (ATP) of body regulates which
pathway gets energy
 Same in ruminants and non-ruminants
Fate of Absorbed Glucose
 1st Priority: glycogen storage
 Stored in muscle and liver
 2nd Priority: provide energy
 Oxidized to ATP
 3rd Priority: stored as fat
 Only excess glucose
 Stored as triglycerides in adipose
Pyruvate Metabolism
 Three fates of pyruvate:

 Conversion to lactate (anaerobic)

 Conversion to alanine (amino acid)
 Entry into the TCA cycle via pyruvate
dehydrogenase pathway (create ATP)
 Production of
blood glucose
Glycogenolysis
 2 hours after a meal
 the primary source of blood
glucose during the first few hours
of fasting
Gluconeogenesis
 after consumption of the liver
glycogen
 lactate (muscle, erythrocytes),
amino acids (muscle), glycerol
(adipose tissue)
Fate of Product of Glycolysis- Pyruvate
- Pyruvate is at a central branch point
in metabolism.
Recall:
Aerobic pathway - through
citric acid cycle and respiration;
this pathway yields far more energy
and will be discussed later.

NADH + O2  NAD+ + energy

Pyruvate + O2  3CO2 + energy
Cori Cycle

Lactate is
converted
to pyruvate
in the liver
Two anerobic pathways:

- to lactate via lactate dehydrogenase

- to ethanol via ethanol dehydrogenase

- Note: both use up NADH produced

so only 2 ATP per glucose consumed
Pyruvate metabolism
 Convert to alanine and export to blood
Glutamate -Ketoglutarate
COO– COO–
C O HC NH 3+
Alanine amino transferase
CH3 (AAT) CH3
Pyruvate Alanine
Keto acid Amino acid
 Pyruvate Dehydrogenase Complex
(PDH)
 Prepares pyruvate to enter the TCA cycle

Aerobic Conditions
Electron TCA
Transport Cycle
1. Lactate Fermentation
Enzyme = Lactate Dehydrogenase

COO- COO-
C=O + NADH + H+  H-C-OH + NAD+
CH3 CH3
pyruvate lactate

-Note: uses up all the NADH(reducing

equivalents) produced in glycolysis.
Helps drive glycolysis by using up
NADH
 reversible so pyruvate can be
regenerated in alternative metabolism
 lactate fermentation important in
red blood cells, parts of the retina,
and in skeletal muscle cells during
strenuous exercise.
-Also important in plants and in
microbes growing in absence of O2.
--
Lactate Dehydrogenase (LDH) has
multiple forms. It is an isozyme.
Two polypeptides M and H come
together to form LDH. It is a tetramer
so a mixture is formed:
M4, M3H, M2H2, MH3 and H4

M M M H H H H H H H
M M M M MM M H H H
 Skeletal muscle and liver contain
predominantly the “M” forms;
heart the “H” forms. During and
after myocardial
infarction (heart
attack), heart
cells die releasing
LDH into the
circulation.

 Diagnostic.
LACTIC ACID (CORI) CYCLE
glucose
glucose glucose
glucose-6-P glucose-6
glycogen glycogen
ATP ATP
NADH Blood NADH
pyruvate pyruvate
lactate lactate
lactate
Liver Muscle

The liver uses most of this lactate to
make glycogen. Only small amounts
of free glucose released.

 Glycogen can be broken down into

glucose when needed.
2. Alcoholic Fermentation

COO- CO2 CH2OH


C=O C + NADH CH3
+CH3OH  CH3 
NAD+

pyruvate acetaldehyde ethanol

 pyruvate decarboxylase- irreversible
 alcohol dehydrogenase- reversible
- pathway is active in yeast.
- second step helps drive glycolysis
-second step is reversible
- reverse is ethanol oxidation,
eventially yields acetate, which
ultimately goes into fat synthesis.
- ethanol  acetaldehyde  acetate
- humans have alcohol dehydrogenase
in liver which mainly disposes of
ethanol.
- acetaldehyde is reactive and toxic.
Summary Glucose
of Reactions 2 ATP
2 NADH
2 pyruvate
2 NADH 2 NADH
anaerobic
anaerobic
2 ethanol + CO2 2 lactate

2 acetyl CoA + 2 CO2

O2 aerobic
4 CO2 + 4 H2O
Siklus 2,3 Biphosphoglicerat
2.3 Biphosphoglycerate(BPG)
Human Hb and binding site for 2,3 BPG
 The rate of Glycolysis will influent the affinity
oxygen and Hemoglobine,with the intermediate
2,3 BPG pathway
 Disorder in glycolysis will influent the affinity
hemoglobine and oxygen.
 Defficiency Piruvat kinase, so the level of 2.3
BPG will increase.
 The affinity of oxygen and hemoglobine loose,
and hypoxia in the tissue
 Anemia hemolytic.
Deficiency Hexokinase
 - Genetic disease

 - 2.3 BPG in RBC low

 - Affinity Hb and Oxygen is very strong
(abnormal)
 - Hypoxia in the tissue
 Defficiency Piruvate kinase
(Anemia Hemolitik)
 - Blockade The end of glycolytic pathway, The
affinity of oxygen and Hb decrease. turun.
 - The production of ATP is not enough, so it
decrease the activity of Na+ & K+, stimulate ion
ATP ase pump.
 It will keep the membran cell of RBC.
 Defficiency Piruvate Kinase will make RBC
Lysis.
The important pathways of glucose metabolism. Note
that the glycogen degradations pathways end in -lysis,
while the glycogen synthesis pathways end with -genesis.
 Glycogen metabolism
Glucose is the main source of energy for brain, cells
without mitochondria (RBC) and essential source of energy
for exercising muscles because it substrate for anaerobic
glycolysis Diet
Source of Blood glucose Glycogen
gluconeogenesis
1) Dietary intake of glucose or glucose precursors as starch,
monosaccharides (Fructose), disaccharides (Sucrose, maltose, lactose)

2) Glycogen: provide a rapid supply for glucose in the absence of dietary

glucose. Glycogen is rapidly degraded into glucose

3) Gluconeogenesis: provide sustained synthesis of glucose stimulated by

low blood glucose (slow)
Structure and function of glycogen
The main store of glycogen in the body is liver (100 g) and skeletal
muscles (400g)
In muscle: glycogen Glucose to produce ATP and energy
In liver: Glycogen Maintain blood glucose
Why NOT store excess glucose as free glucose instead of glycogen?

Fluctuation of glycogen stores

-Liver glycogen stores increase during the well-fed state
and are depleted during a fast
-Muscle glycogen is NOT affected by short periods of
fasting and is moderately decreased in prolonged fasting
while it is affected by exercise.
Metabolism of
glycogen
Synthesis of Glycogen(Glycogenesis)
α-D- glucose is the monomer
 Occurs in the cytosol
 Requires energy supplied by ATP and UTP

α-D- glucose attached

to UDP (glucose-UDP) is
the source of all the
glycosyl residues that
are added to the growing
glycogen
 Glycogenesis
 Glycogen synthesis
 Occurs in cytosol of liver,muscle& kidney
 Occurs when blood glucose levels are high
 Excess glucose is stored (limited capacity)
 liver and muscle are major glycogen storage sites
 liver glycogen used to regulate blood glucose levels
 brain cells cannot live for > 5 minutes without glucose
 muscle glycogen used to fuel an active muscle
GLIKOGENESIS
 Glycogen phosphorylase of liver
as a glucose sensor.

1. In liver, when blood glucose levels return to normal, glucose enters

hepatocytes and binds to an inhibitory allosteric site on phophorylase
a.
2. This binding produces a conformational change that exposes the
phosphorylated Ser residues to PP1
3. And then dephosphorylated and inactivated.
Glycogen synthase is regulated by
phosphorylation &dephosphorylation
Insulin Regulation On Muscle Cells

Problem is all about cytoplasmic concentration of glucose

 Glycogen Synthesis
Glucose units are activated for transfer by formation
of sugar nucleotides
 What are other examples of "activation"?

 acetyl-CoA, biotin, THF,

 Leloir showed in the 1950s that glycogen
synthesis depends on sugar nucleotides
 UDP-glucose pyrophosphorylase
 a phosphoanhydride exchange
 driven by pyrophosphate hydrolysis
UDP-glucose synthesis
UDP-glucose is the source of
glucosyl groups that used in
glycogen synthesis
Glycogen synthase is
UDP + ATP nucleoside diphosphate kinase UTP + ADP
responsible for making α-
1  4 linkage. It can only
elongate existing chains
and cannot initiate chain
synthesis.
* Fragment of glycogen
can serve as a primer

Elongation of glycogen chains

by Glycogen Synthase
Involve transfer of glucose
units from UDP-glucose to the
non-reducing end of the
growing chain forming α-14
glycosidic linkage (anomeric
hydroxyl of carbon 1 of the
activated glucose and carbon
4 of accepting glucose
residue).
 * Formation of branches in glycogen
* Glycogen is highly branched molecule (every 8 residues)
 increase solubility and size compared to the non-
branched amylose.
* Branching increases the number of non-reducing ends to
which glucosyl residues can be added or removed 
accelerate the rate of glycogen synthesis or degradation.
* Synthesis of branches by glycosyl 4:6 transferase
Branches are made by “branching enzyme” that called
Amylo-(α14)(α16)transglycosylase or Glucosyl(4:6)
transferrase This enzyme transfers 5-8 glycosyl
residues from the non-reducing end to another residue
forming α-1 6 linkage. Old and new non-reducing ends
are available to be further elongated by Glycogen
 The OH gp of specific
tyrosine side chain is
the initial site of
attachment
* Fragment of
glycogen can serve
as a primer
* Glycogen initiator
In absence of synthase: transfer the
glycogen, specific first molecule of glucose
from UDP-glucose to
protein glycogenin glycogenin.
can serve as an Then additional glucose
unit is transferred to
acceptor of form short chain
glucose residues.
 Glycogen Synthase
Forms -(1 4) glycosidic bonds in glycogen
 Glycogenin (a protein!) forms the core of a
glycogen particle
 First glucose is linked to a tyrosine -OH
 Glycogen synthase transfers glucosyl units from
UDP-glucose to C-4 hydroxyl at a nonreducing
end of a glycogen strand.
 Note another oxonium ion intermediate
 Control of Glycogen Metabolism
A highly regulated process, involving reciprocal
control of glycogen phosphorylase and glycogen
synthase
 GP allosterically activated by AMP and inhibited
by ATP, glucose-6-P and caffeine
 GS is stimulated by glucose-6-P

 Both enzymes are regulated by covalent

modification - phosphorylation
Phosphorylation of GP and GS
Covalent control
 Edwin Krebs and Edmond Fisher showed in 1956
that a "converting enzyme" converted
phosphorylase b to phosphorylase a(P)
 Phosphorylation causes the amino terminus of the
protein (res 10-22) to swing through 120
degrees, moving into the subunit interface and
moving Ser-14 by more than 3.6 nm
 Nine Ser residues on GS are phosphorylated!
Enzyme Cascades and GP/GS
Hormonal regulation
 Hormones (glucagon, epinephrine) activate
adenylyl cyclase
 cAMP activates kinases and phosphatases that
control the phosphorylation of GP and GS
 GTP-binding proteins (G proteins) mediate
the communication between hormone receptor
and adenylyl cyclase
Hormonal Regulation
of Glycogen Synthesis and Degradation
 Insulin is secreted from the pancreas (to liver)
in response to an increase in blood glucose
 Note that the portal vein is the only vein in the
body that feeds an organ!
 Insulin stimulates glycogen synthesis and
inhibits glycogen breakdown
 Note other effects of insulin
Regulation of liver &muscle glycogen
metabolism:
State Regulators Response

Liver

Fasting Glucagon ↑, Insulin ↓ Glycogen degradation ↑

cAMP ↑ Glycogen synthesis ↓
Carbohydrate meal Glu ↑, Glucagon ↓, Insulin ↑ Glycogen degradation ↓
cAMP ↓ Glycogen synthesis ↑
Exercise and stress Adrenalin ↑ Glycogen degradation ↑
cAMP ↑, Ca2+-calmodulin ↑ Glycogen synthesis ↓

Muscle
Fasting (rest) Insulin ↓ Glycogen synthesis ↓
Glucose transport ↓
Carbohydrate meal (rest) Insulin ↑ Glycogen synthesis ↑
Glucose transport ↑
Exercise Epinephrine ↑ Glycogen synthesis ↓
AMP ↑, Ca2+-calmodulin ↑, Glycogen degradation ↑
cAMP ↑ Glycolysis ↑
Glucose homeostasis:
 maintenance of blood glucose levels near 80 to 100 mg/dL (4,4-5,6
mmol/l)
 insulin and glucagon (regulate fuel mobilization and storage)

Hypoglycemia prevention:
1. release of glucose from the large glycogen stores in the liver
(glycogenolysis)
2. synthesis of glucose from lactate, glycerol, and amino acids in liver
(gluconeogenesis)
3. release of fatty acids from adipose tissue (lipolysis)

Hyperglycemia prevention:
1. conversion of glucose to glycogen (glycogen synthesis)
2. conversion of glucose to triacylglycerols in liver and adipose tissue
(lipogenesis)
Pathways regulated by the release
of:
 glucagon (in response to a lowering of blood glucose levels)
 insulin (in response to an elevation of blood glucose levels)
 Major sites of insulin action on fuel
metabolism:
The storage of nutriens
• glucose transport into
muscle and adipose tissue
• glucose storage as
glycogen (liver, muscle)
• conversion of glucose to
TG (liver) and their storage
(adipose tissue)
• protein synthesis (liver,
muscle)
• inhibition of fuel
 Major sites of glucagone action on
fuel metabolism:
Mobilization of energy
stores
1. release of glucose from
liver glycogen
2. stimulating
gluconeogenesis from
lactate, glycerol, and
amino acids (liver)
3. mobilizing fatty acids
(adipose tissue)
Hormonal Regulation II
Glucagon and epinephrine
 Glucagon and epinephrine stimulate glycogen
breakdown - opposite effect of insulin!
 Glucagon (29 res) is also secreted by pancreas
 Glucagon acts in liver and adipose tissue only!
 Epinephrine (adrenaline) is released from adrenal
glands
 Epinephrine acts on liver and muscles
 The phosphorylase cascade amplifies the signal!
 CH2OH CH2OH
O O
O O O -[1-6] linkage

CH2OH CH2OH CH2 CH2OH

........ O O O O
O O O O

-[1- 4] linkages

. The glycogen structure showing the glycosidic bonds

 Glycogenesis
 Liver
 7–10% of wet weight
 Use glycogen to export glucose to the bloodstream when
blood sugar is low
 Glycogen stores are depleted after proximately 24 hrs of
fasting (in humans)
 De novo synthesis of glucose for glycogen
 Skeletal muscle
 1% of wet weight
 More muscle than liver, therefore more glycogen in muscle, overall
 Use glycogen (i.e., glucose) for energy only (no export of
glucose to blood)
 Use already-made glucose for synthesis of glycogen
 Glucose
ATP
Hexokinase
(muscle)
Glucokinase (Glucose)n (Glucose)n+1
ADP
(liver)
Glucose-6-phosphate
Phospho- Glucose-1-P
glucomutase Uridyltransferase
Glucose-1-phosphate UDP-glucose UDP
Glycogen Synthase
UTP PPi

Pathway of glycogen synthesis (glycogenesis).

 Control of enzyme activity

Rate limiting step

Glycogen synthesis
Glucose 6-P→ glucose 1-P.
glucose 1-P + UTP→UDP-glucose + PPi.
PPi + H2O→ 2 Pi.
UDP-glucose + glycogenn → glycogenn+1.
UDP + ATP → UTP + ADP.

Glucose 6-P + ATP + glycogenn + H2O →

glycogenn+1 + ADP + 2Pi.

Only one ATP is used to store one glucose

residue in glycogen.

(nucleoside diphosphokinase)
Glycogen synthesis and breakdown
are reciprocally regulated
Red=inactive forms,
green = active forms.

Active Inactive

Protein phosphatase 1 (PP1) regulates

glycogen metabolism.
Glycogenolysis
 Glycogen degradation
 Occurs in cytosol
 Signal that glucose is needed is given by
hormones
 epinephrine stimulates glycogen breakdown in
muscle
 glucagon which stimulates glycogen breakdown
in liver in response to low BG
 used to sustain blood glucose level between meals
and to provide energy during an
emergency/exercise
 Glycogen Catabolism
Getting glucose from storage (or diet)
 -Amylase is an endoglycosidase
 It cleaves amylopectin or glycogen to maltose,
maltotriose and other small oligosaccharides
 It is active on either side of a branch point, but
activity is reduced near the branch points
 Debranching enzyme cleaves "limit dextrins"
 Note the 2 activities of the debranching enzyme
 Glycogen
Pi
glycogen
phosphorylase LIVER PATHWAY
Glucose-1-phosphate
phosphoglucomutase
glucose-6-phosphatase
Glucose-6-phosphate Glucose

glycolysis Pi
(inhibited by lack of X
fructose-2,6-bisP

Glycogenolysis and the fate of glycogen in liver and kidney

 Glycogen MUSCLE PATHWAY
Pi
glycogen
phosphorylase
Glucose-1-phosphate
phosphoglucomutase

Glucose-6-phosphate

glycolysis anaerobic
Pyruvate Lactate
pyruvate lactate dehydrogenase
dehydrogenase
Acetyl CoA CO2
citric acid cycle
aerobic

. Glycogenolysis and the fate of glycogen in muscle.

Glikogenesis & Glikogenolisis
 Glucose anabolism
 Glucose storage:
glycogenesis
 glycogen formation is
stimulated by insulin
 glucose not needed
immediately is stored
in the liver (25%) and
in skeletal muscle
(75%)
 Glucose release:
glycogenolysis
 converts glycogen to
glucose
 occurs between
meals, stimulated by
glucagon and
epinephrine
 SIMPLISTIC SUMMARY:
-- Epinephrine(adrenaline) and glucagon
glycogenolysis & inhibit glycogenesis
via a cAMP and a phosphorylation
cascade.  release glucose
-- Glycogenesis is stimulated by
insulin in a pathway ending in the
dephosphorylation of glycogen
synthase.
-- Glycogenolysis is also inhibited
via dephosphorylation.
 take up glucose
 Glycogen Storage Diseases:
A family of serious, although not
necessarily fatal, diseases caused by
mutations in the enzymes involving
in glycogen storage and breakdown.
 Glycogen Storage Diseases

Type I: Von Gierke Disease; Glucose-6-phosphatase Defect

 Hypoglycemia occurs due to defect of the final step of gluconeogenesis.

 This disease, affects only liver and renal tubule cells
 Decreased mobilization of glycogen produces hepatomegaly.
 Decreased gluconeogenesis causes increased lactate leading to lactic acidemia.

Type V: McArdle Disease; Skeletal Muscle Glycogen Phosphorylase Defect

 Skeletal muscle is affected, whereas the liver enzyme is normal.

 Temporary weakness and cramping of skeletal muscle occurs after exercise.
 There is no rise in blood lactate during strenuous exercise.
 Muscle contains a high level of glycogen with normal structure

Type VI: Hers Disease; Liver Glycogen Phosphorylase Defect

 Liver is affected, whereas the skeletal muscle enzyme is normal.

 Marked hepatomegaly occurs due to a high level of glycogen with normal structure..
 Following administration of glucagon, there is no increase in blood glucose.
Pentose Phosphate Pathway=
Hexose Monophosphat Shunt
Generation of NADPH and Pentoses
Has 2 functions
1.Generate reducing equivalents NADPH (reduced cosubstrate/
coenzyme) needed for fatty acid synthesis, folate reduction
2. Produce ribose 5-phosphate needed for DNA and RNA
synthesis

Occurs in cytosol of cells particularly important in anabolic

tissues,liver, adrenal cortex, mammary glands and fat tissues
muscle cells do NOT have HMS enzymes
Pentose Phosphate Pathway
Glucose-6- 6-Phospho- 6-Phospho-
phosphate glucono- gluconate
lactone

D-Ribulose-
5-phosphate

D-Ribose-
RNA or DNA 5-phosphate
 Oxidative branch

Glucose-6-P-dehydrogenase
ATP ADP NADP NADPH
6-Phosphogluconate
Glucose Glucose 6-P
NADP
6-Pgluconate dehydrogenase
Ribulose 5-P NADPH
CO2
Non-oxidative branch

Xylulose 5-P Ribose 5-P (5 carbons)

Transketolase

Glyceraldehyde 3-P Transketolase TDP

Glyceraldehyde 3-P Sedoheptulose 7-P (7 carbons)
TDP
Fructose 6-P Transaldolase
Erythrose 4-P Fructose 6-P

NADPH is used for biosynthetic reactions and glutathione metabolism

Glyceraldehyde-3-P and fructose-6-P return to the glycolytic pathway

A scenario in which the cell requires NADPH but does not require ribose-5-P
 Nucleic acids
Ribulose 5-P

Xylulose 5-P Ribose 5-P (5 carbons)

Glyceraldehyde 3-P Transketolase
Transketolase TDP
Glyceraldehyde 3-P Sedoheptulose 7-P (7 carbons)
TDP
Fructose 6-P Transaldolase
Erythrose 4-P Fructose 6-P

Ribose-5-P is the sugar required for the synthesis of nucleic acids

Oxidative branch is feedback inhibited by excess NADPH at glucose-6-P
dehydrogenase
A scenario in which the cell requires ribose-5-P but does not require NADPH
 Glucose-6-P-dehydrogenase
ATP ADP NADP NADPH

Glucose Glucose 6-P 6-Phosphogluconate

NADP
6-Pgluconate dehydrogenase
Ribulose 5-P NADPH
CO2

Ribose 5-P (5 carbons)

Nucleic acids

A scenario in which the cell requires both NADPH and ribose-5-P

Overview
 Function
 NADPH production
 Reducing power
carrier
 Synthetic pathways
 Role as cellular
antioxidants
 Ribose synthesis
 Nucleic acids and
nucleotides
Characteristics: Tissue Distribution
 Demand for NADPH
 Biosynthetic pathways
 FA synthesis (liver, adipose, mammary)
 Cholesterol synthesis (liver)
 Steroid hormone synthesis (adrenal, ovaries, testes)
 Detoxification (Cytochrome P-450 System) – liver
 Reduced glutathione as an antioxidant (RBC)
 Generation of superoxide (neutrophils)
Characteristics:
Oxidative and Non-oxidative Phases
 Oxidative phases
 Reactions producing
NADPH
 Irreversible
 Non-oxidative phases
 Produces ribose-5-P
 Reversible reactions feed
to glycolysis
NADPH producing reactions
 Glucose-6-P dehydrogenase
 6-P-gluconate dehydrogenase
The Pentose Phosphate Pathway:
Non-oxidative phases
Regulation
 Glucose-6-P dehydrogenase
 First step
 Rate limiting
 Allosteric Regulation
 Feedback inhibited by NADPH
 Inducible enzyme
 Induced by insulin
HMS ( Hexose Monophospat Shunt)
 Nicotinamide adenine dinucleotide phosphate
 phosphorylated form of reduced nicotinamide
adenine dinucleotide (NADH)
 generated in a series of reactions comprising
the oxidation-reduction phase of HMS
 Ribose 5-phosphate
 sugar used as the backbone of DNA and RNA
 Cell’s requirement for ATP (glycolysis) or
NADPH and ribose 5-P (HMS) determines which
path it will take.
Stages of HMS
 Reactions occur in 3 main stages
 oxidation-reduction
 generation of NADPH
 isomerization stage
 generation of ribose 5-phosphate
 carbon bond cleavage-rearrangement stage
 conversion of three 5-carbon sugars to two 6-
carbon sugars (Fructose 6-phosphate) and one 3-
carbon sugar (Glyceraldehyde 3-phosphate)
 these series of reactions occur in cells where
demand for NADPH is high
 F 6 P can be converted back to G 6 P which can re-enter
HMS
Reactions of Stages 1 and 2
 G6P is oxidized to 6-phosphoglucono-lactone by G6P
dehydrogenase that uses NADP as coenzyme
 produces NADPH and 6-phosphoglucono
 6-phosphoglucono is hydrolyzed (addition of water) to
6-phosphogluconate
 6-phosphogluconate is oxidized by 6 phosphogluconate
dehydrogenase
 produces NADPH and ribulose 5 phosphate
 Ribulose 5-phosphate is isomerized to ribose 5
phosphate
Regulation of Metabolism Revisited
 Allosteric Enzyme Modulation
 enzymes can be stimulated or inhibited by certain
compounds
 modulators act by altering conformational
structure of their allosteric enzymes
 causes shifts between relaxed and tight conformations
 relaxed is most active
 ratio of ADP (or AMP) to ATP is important in
regulation of energy metabolism
Allosteric Enzyme Modulation
 low ADP:ATP ratio signals less need to
produce ATP
 inhibition of key enzymes in glycolysis and the
TCA cycle
 PFK, PDH, CS, and isocitrate dehydrogenase
 high ADP:ATP ratio signals need for ATP
 activation of the above enzymes
 ATP is end product in oxidative catabolism and its
accumulation would signal to decrease catabolic
pathway activity
Allosteric Enzyme Modulation
 ratio of NADH to NAD+ is also important in
regulation
 NADH is end product of catabolic pathway
 accumulation would signal to decrease activity
 diminution would signal to increase activity
 key enzymes are affected in glycolytic and TCA
cycle
 PK, PDH, CS, isocitrate dehydrogenase and alpha KG
dehydrogenase
Role of NADPH in the RBC
 Production of superoxide
 Hb-Fe2+-O2 -> Hb-Fe3+ + O2-.
 Spontaneous rxn, 1% per hour
 O2-. + 2H2O -> 2H2O2
 Both O2-. & H2O2 can produce reactive free
radical species, damage cell membranes, and
cause hemolysis
Pentose Phosphate Pathway
Glucose 6-phosphate dehydrogenase deficiency
Detoxification of Superoxide Anion
and Hydrogen Peroxide
 Antioxidant enzymes
 Superoxide dismutase
 Glutathione peroxidase
 Glutathione reductase
Repetition:

1. 3 key enzymes for the regulation of glycolysis (their

activation). The role of Fructose 2,6-P in the regulation of
glycolysis and gluconeogenesis.
2. 3 key sites for the regulation of gluconeogenesis (their
activation).
3. The signal pathway for the activation of glycogen
degradation by glucagon.
4. Main regulators of glycogen degradation in liver and
muscle.
5. Pathways preventing hypoglycemia and hyperglycemia.
Pictures used in the presentation:
Marks´ Basic Medical Biochemistry, A
Clinical Approach, third edition, 2009 (M.
Lieberman, A.D. Marks)
References for Further Readings:
Latest editions of:
•Biochemistry by L. Stryer, Freeman & Company New York ….
•Harper’s Biochemistry by R.K. Murray, D.K. Granner, P.A. Mayes &
V.W. Rodwell. Biochemistry. Appleton & Lange, New York, Connecticut,
California.
•Biochemistry by T. Mckee & J. Mckee, Wm. C. Brown Publishers,
London, Madrid, Tokyo ….
•Biochemistry by I.D.K. Halkerston, Second Edition, John Wiley & Sons
Inc., Williams E Wilkins, USA.
• Textbook of Biochemistry With clinical Correlations by T.M. Devlin,
Wiley-Liss Publication, New York, Toronto ….
•Biochemistry Illustrated by P.N. Campbell & A.D. Smith, Longman
Group UK Ltd, UK.
•Principles Of Biochemistry byDr.G.L.
27 August 2019
Zubay, W.W. Parson & D.E. Vance,
Mohamed Z Gad 136
Wm. C. Brown Publishers, Iowa, Melbourne, Ox for d.