1

CARBOHYDERATE METABOLISM

Dr GULSHAN ABBAS
PROFESSOR
Dept of Biochemistry & Molecular
Biology
Hitec IMC
• "And from the fruits of date palm and grapes you
get wholesome drink and nutrition: Behold in this is
a sign for those who are wise" (Quran 16:67)

• "But waste not by excess for God loves not the

wasters" (Quran 7:31)
• "And if God touches thee with affliction, none can
remove it but He: if He touches thee with
happiness He has power over all things." (Quran
6:17).
 b
What is metabolism?
– Organized reactions by enzymes in multiple
steps result into pathways
– Different pathways interact with each other in
a cell, a tissue or the body
– Catabolic pathways (degradative)
– Anabolic pathways (constructive)

Metabolic map
– Each pathway required for metabolism of
carbohydrates, proteins and lipids is part of
metabolic map
Metabolic map
Overview of intermediary metabolism

Introduction

1. Anabolic pathways
2. Catabolic pathways
3. Amphibolic pathways
The basic metabolic pathways process the
major products of digestion

Glucose, fatty acids and amino acids, glycerol

All these are converted to common product i.e.
acetyl CoA
Overview of CHO Metabolism
Lipid metabolism is concerned mainly
with fatty acids and cholesterol

–Produces acetyl CoA in high quantity

via -oxidation and more ATP
–Acetyl CoA for cholesterol synthesis
–Acetyl CoA for ketogenesis
 PROTEIN METABOLISM
Much of amino acid metabolism involves
transamination

– Protein synthesis
– Transamination
– Deamination
– Urea synthesis
 REGULATION OF METABOLISM
Metabolic pathways in all cells, tissues and organs
are coordinated by a communication system
– Hormones
– Neurotrasmmitters
– Availability of nutrients
– Energy states of body
– Different conditions of body
 REGULATION OF METABOLISM (Contd)

A. Intracellular signals (rapid response)

– Availability of substrates
– Product inhibition
– Levels of allosteric activators or inhibitors

B. Communication between cells (intercellular)

– Long term integration of metabolism
– Chemical signaling between cells by hormones and
neurotrammitters
 REGULATION OF METABOLISM (Contd)

C. Second messenger system

– Hormone acts on receptor (signal detector)
– Second messenger translates hormone or
neurotransmitter binding into cellular
response
• Calcium / phosphatidylinositol system
• Adenylyl cyclase system
Structure of a typical memberane receptor
REGULATION OF METABOLISM (Contd)

D. Adenylyl cyclase system (via G-proteins)

• Membrane bound receptors

• Chemical signals (hormones and

neurotransmitters) bind receptor

• Cyclic AMP second messenger produced from

ATP by Adenyle cyclase acting through a cascade
of events
1. GTP dependent regulatory protein (G-proteins)

– Trimeric G-proteins ie →-subunit and  dimer

2. Protein kinases

– Cyclic AMP activates protein kinases A &B

– Protein kinases phosphorylate some proteins that

act directly on channels or become activated or
inhibited (mainly enzymes)

– Protein kinase - C is not cyclic AMP dependent

 Characteristics of G proteins
1. G protein is an  trimeric protein which binds guanine nucleotides.
2. They function to couple integral membrane receptors to target
membrane-bound enzymes.
3. They can be considered molecular switches wherein…
GDP (inactive) → GTP (active) + 
4. The dissociated  subunit expresses GTPase activity.

5. GTPS blocks GTPase

activity of GTP.
 REGULATION OF METABOLISM (Contd)

3. Dephosphorylation of proteins
Protein phosphatase removes phosphates
and inactivates kinases
4. Hydrolysis of cAMP by c-AMP
phosphodiestrase
This enzyme is inhibited by methylxanthine
derivatives eg theophylline and ceffiene
Actions of CAMP
B. The second messenger Is
cGMP

• Atrial natriuretic factor (ANF)

• Nitric oxide (NO)

C. The second messenger Is calcium or

phosphatldyllnositols (or both):

• Acetylcholine (muscarinic)
• a-Adrenergic catecholamines
• Cholecystokinin
• Gastrin
• Gonadotropin-releasing hormone (GnRH)
• Oxytocin
• Thyrotropin-releasing hormone (TRH)
D.The second messenger Is a kinase
or phosphatase cascade
• Insulin
• Prolactin (PRL)
• Erythropoietin (EPO)
• Growth hormone (GH)
• Chorionic somatomammotropin (CS)
• Epidermal growth factor (EGF)
• Fibroblast growth factor (FGP)
• Insulin-like growth factors (IGF-I, IGF-II)
• Nerve growth factor (NGF)
• Platelet-derived growth factor (PDGF)
 The second messenger Is a
kinase or phosphatase cascade
 The second messenger Is a
kinase or phosphatase cascade
 Biomedical importance of
Metabolism
To understand abnormal metabolism in
diseases e.g. nutritional deficiency,
enzyme deficiency, abnormal secretion
of hormones, metabolic diseases like
diabetes mellitus, hyperlipidemias and
glycogen storage diseases.
 Carbohydrate
Metabolism
Digestion of carbohydrates

Absorption of carbohydrates
Sodium dependent transport

Sodium independent transport

 FORMS IN WHICH INGESTED
1. Polysaccharides :-
➢ Starch
➢ Glycogen
2. Disaccharides :-
➢ Lactose
➢ Sucrose
3. Monosaccharides :-
➢ Glucose
➢ Fructose
Digestion of Carbohyderates
 ABSORPTION
➢Transport of the products of digestion from
the intestinal tract into the blood
►The final products of carbohydrate digestion
in the alimentary tract are almost entirely
glucose, fructose, and galactose

► Glucose representing, on average, about 80

per cent of these

►After absorption from the intestinal tract,

much of the fructose and almost all the
galactose are rapidly converted into glucose in
the liver
Glucose thus becomes the final common

pathway for the transport of almost all

carbohydrates to the tissue cells

 TRANSPORT OF GLUCOSE
INTO CELLS
Plasma glucose can not diffuse directly into cells
Enters by one of the two transport mechanisms

A. Na+ independent facilitated diffusion

transport
• Transported by glucose transporters
• Isoform GLUT-1 to GLUT-14
• These transporters are present in cell
membranes
Tissue specificity of GLUT gene expression
 GLUCOSE TRANSPORTERS
►GLUT-1 (RBCs & Brain)
►GLUT-2 (Liver, kidneys, β cells)
►GLUT-3 (Neurons)
►GLUT-4 (Adip. tis & sk. muscle)
►GLUT-5 (Transport of fructose)
►GLUT-7 (Across ER)
►SGLUT-1 (Intest,inhi by ouabain)
►SGLUT-2 (PCT kidney)
Transport of Glucose into the cell
 TRANSPORT OF GLUCOSE
INTO CELLS
B. Na+ monosaccharide cotransporter system

– Energy (ATP) dependent

– Against Concentration gradient to cells

– Carrier mediated process coupled with

Na+

– In intestine, renal tubules and choroid

plexus
SGLT
 CARBOHYDRATE METABOLISM
Catabolic pathways (convergent process)
– Capture chemical energy in the form of ATP, from
energy rich fuel molecules
– Energy generation by degradation of carbohydrates,
fats and proteins in three stages.
1. Hydrolysis of complex molecules to building blocks
2. Conversion of building blocks to simple
intermediates Acetyle CoA along with small ATP
3. Oxidation of acetyl CoA
– Via TCA cycle
– Large ATP via FADH and NADH
 CARBOHYDRATE METABOLISM

Anabolic pathways (divergent process)

– Small molecules to complex molecules

– Reduction occurs by NADPH
– Requires ATP
– Divergent process
 PATHWAYS INVOLVED IN CARBOHYDRATE
METABOLISM
• Glycolysis
• Tricarboxylic acid cycle (TCA cycle) Krebs citric
acid cycle)
• Glycogen metabolism
• Gluconeogenesis
• Metabolism of monosaccharides and
disaccharides
(fructose and galactose)
• Hexose monophosphate shunt (HMP)
 GLYCOLYSIS
►Main metabolic pathway for glucose
►Found in cytosol of all tissues
►Aerobically and anaerobically
►RBCs totally rely on it for energy
►(Beyond pyruvate, mitochondria and O2
both required)
 CLINICAL IMPLICATIONS

►Heart muscle ischemia

►Fatigue (phosphofructokinase)

►Hemolytic anemias (Pyruvate kinase)

►Lactic Acidosis (Pyruvate dehydrogenase)

 GLYCOLYSIS
(Embden Meyerhof Pathway)
►Glycolysis is a pathway for the breakdown of
glucose to provide energy to cells
►Pyruvate is the end-product in cells with
mitochondria and lactate is the end-product
in cells without them
– Aerobic Glycolysis
– Anaerobic glycolysis
 GLYCOLYSIS
TWO STAGES
• Stage-1 Energy [ATP] investment phase

• Stage-II Energy [ATP] generation phase, at

substrate level (2 ATP) [Anaerobic]
8 ATP (aerobic]
Two phases of Aerobic Glycolysis
 GLYCOLYSIS
►Stage 1 (Preparatory): 1 molecule of glucose
2 molecules of glyceraldehyde 3-
phosphate
►Stage 2 (Payoff): 2 molecules of glyceraldehyde 3-
phosphate 2 molecules of pyruvate

AND
4 ADP 4ATP
 REACTIONS / STEPS OF GLYCOLYSIS
Step 1
1. Phosphorylation of glucose to glucose – 6
phosphate
– Glucose can not come out of cell and is trapped
– This is irreversible and committed step
– This is the cross road of metabolism of glucose
– Enzymes are hexokinase and glucokinase
• Fate of glucose-6 phosphate
• Glycolysis to pyruvate / lactate
• Glucose (gluconeogenesis)
• Glycogen synthesis
• Hexose monophosphate pathway
Energy investment phase, Phosphorylation of Glucose
 REACTIONS / STEPS OF GLYCOLYSIS

A. Hexokinase 1-3
– Present in most tissues
– Regulatory enzyme of glycolysis
– Broad substrate specificity
– Inhibited by reaction product – G. 6.
phosphate
– Has low km (high affinity) and can use
glucose at low concentration
– Has a low Vmax and cannot
phosphorylate more sugars than the cell
can use
 B. Glucokinase (Hexokinase 4)
– Present in liver parenchymal cells and islets of the
pancreas
– Main enzyme responsible for phosphorylation of
glucose in hyperglycemia (fed state)
– Acts as the glucose sensor in -cells of pancreas
Kinetics
– Much higher Km for glucose and high V max
– Active only in hyperglycemia after meal and
removes the flood of glucose by the liver
– GLUT-2 equilibrate the blood glucose across
the hepatocyte membrane rapidly
Regulation by fructose-6 phosphate and glucose
» Not inhibited by glucose-6 phosphate
» Indirectly inhibited by F-6 phosphate
» Stimulated indirectly by glucose
» Glucokinase regulatory protein present in the
nucleus of hepatocytes regulates this process
Regulation by insulin
» Increased glucose level increases insulin by  cells
» Insulin increase activity of glucokinase
» Insulin promotes transcription of glucokinase gene
» Diabetes mellitus due to deficiency in hepatic
glucokinase
Regulation of Glucokinase by Glucokinase regulatory protein
 HEXOKINASE GLUCOKINASE (liver)

Low Km (high affinity) for High Km (low affinity) for

glucose, low Vmax glucose, high Vmax

Inhibited by glucose 6- Inhibited by fructose 6-

phosphate phosphate

Found in all tissues Absent in muscles

Not specific Deficient in diabetes

 Step 2

ISOMERIZATION OF GLUCOSE – 6
PHOSPHATE TO FRUCTOSE – 6 –
PHOSPHATE

Enzyme : Phosphoglucose isomerase

Isomerisation of Glucose-6-Phosphate to Fructose-6 phosphate
 Step 3
Phosphorylation of fructose – 6 phosphate
to from fructose –1,6 - bisphosphate

• Enzyme : phosphofructokinase –1 (PFK-1)

• Rate limiting step of gluconeogenesis
• Most important control point
• PFK –1 is regulated by a complex
mechanism
1. Regulation by energy levels within the cell,
allosterically by following
a. ATP – (energy signal) inhibits PFK-1
b. Citrate inhibits PFK-1
c. AMP stimulates PFK-1

2. Regulation by fructose 2, 6 bisphosphate

a. Most potent activator of PFK-1
b. This acts as an intracellular signal
c. This is formed by the action of enzyme
phosphofructokinase 2 (PFK-2)
 3. Hormonal regulation of F-2, 6-B phosphate
formation
a. During the well fed state (insulin increases)
– More insulin more F 2,6, bisphosphate due to
increased PFK-2
– Decrease level of glucagon less active FBP-2
– More glycolysis and less gluconeogenesis

b. During starvation (fasting)

– Low insulin, more glucagon, low PFK-2 low F 2, 6
bisphosphate
– More active fructose bisphosphatase-2(FBP-2)
– Less glycolysis and more gluconeogenesis
Actions of CAMP
 Step 4
Cleavage of fructose 1, 6-bisphosphate to
glyceraldehyde–3 phosphate and dihydroxy
acetone phosphate (DAP)

Enzyme =Aldolase – A

(Aldolase – B, in the liver and kidney cleaves

Fructose, 1 phosphate to Glyceraldehyde
and dihydroxyacetone phosphate)
This step represents cleavage of a hexose into two
trioses.
Only glyceraldehyde 3-phosphate is on the direct
pathway of glycolysis
1
1
2
2
3 4
3
4 5
5 6
6
 Step 5

ISOMERIZATION OF
DIHYDROXYACETONEPHOSPHATE TO
GLYCERALDEHYDE 3 - PHOSPHATE

Enzymes = Trios phosphate isomerase

Two net Glyceraldehyde molecules are produced

 Isomerization of 3-carbon phosphorylated sugars

1 4

2 5

3 6
 Additional Points
► At equilibrium 96% of triose phosphate is
dihydroxyacetone phosphate
► Net result: Dihydroxyacetone phosphate is funneled
into the main glycolytic pathway
► 2 molecules of glyceraldehyde 3-phosphate are
formed from 1 molecule of fructose 1,6-bisphosphate
► Triose phosphate isomerase accelerates isomerization
by a factor of 1010. Only limited by diffusion of the
substrate into the active site
► Considered a kinetically perfect enzyme
 Step 6
OXIDATION OF GLYCERALDEHYDE –3- PHOSPHATE TO
PRODUCE 1,3 – BISPHOSPHOGLYCERATE (1,3 BPG)

Enzyme = Glyceraldehyde –3- phosphate

Dehydrogenase
Coenzyme = NAD + Pi
NADH + H+ is produced, which is oxidized by two
mechanisms
(a). In Anaerobic condition NADH – linked
conversion of Pyruvate to lactate
(b). Oxidation of NADH via respiratory chain to
produce 3 – ATP
Synthesis of 2,3-bisphosphoglycerate in
red blood cells (2,3 BPG)
Enzyme = Bisphosphoglycerate mutase

This is a shunt reaction in erythrocytes.

2,3 BPG, is very important regulator of O2
delivery by Hb
 The first of the two
Aldehyde
group

energy-conserving
1
2
3
reactions of glycolysis that
NAD+-
will ultimately yield ATP
dependent

1
Mixed
anhydride
Note: The mixed
2
3
anhydride
has a very high
free energy
of hydrolysis.
 Step 7
SYNTHESIS OF 3-PHOSPHOGLYCERATE PRODUCING ATP
FROM 1,3 BISPHOSPHOGLYCERATE

Enzyme = Phosphoglycerate Kinase

1 x ATP is produced at substrate
level (actually 2 ATP)
1
2
ATP is formed as the phosporyl
3

gp on the carboxyl group of

1,3-BPG is transferred to ADP

1
2
3
 Step 8
FORMATION OF 2-PHOSPHOGLYCERATE FROM 3-
PHOSPHOGLYCERATE BY SHIFTING OF PHOSPHATE
GROUP

Enzyme = Phosphoglycerate mutase

 The phosphoryl group is shifted from the C-3 to the C-2
position of glycerate
• Catalyzed by phosphoglycerate mutase

1
1

2
2

3 3
 Step 9
FORMATION OF PHOSPHOENOLPYRUVATE
(PEP)

Enzyme = Enolase
One molecule of H2O is removed from 2-
Phosphoglycerate to form phosphoenolypruvate
(PEP) which contains high energy enolphosphate
 A dehydration reaction in which water is reversibly
removed from 2-phosphoglycerate to from
phosphoenolpyruvate
►Catalyzed by enolase

1 1

2
2
3

3
 Step 10
FORMATION OF PYRUVATE PRODUCING ATP

➢Irreversible reaction
➢Rate limiting step
➢ATP is produced at substrate
level
Enzyme =Pyruvate Kinase
 Transfer of a phosphoryl group from PEP to ADP
►Catalyzed by pyruvate kinase
►Irreversible; An important site of regulation in the liver.
►The second “substrate level phosphorylation”
 Regulation of Pyruvate Kinase
activity

A. Feed Forward regulation, as it is activated

by Fructose 1,6 Bisphosphate
B. Covalent modification by cyclic AMP-
dependent protein Kinase
Glucagon increases phosphorylation of
Pyruvate Kinase and inactivates it.
Pyruvate is now diverted to form
oxaloacetate and then to PEP & glucose
Covalent modification of Pyruvate Kinase results in inactivation of enzyme
 PYRUVATE KINASE DEFICIENCY AND HEMOLYTIC
ANEMIA

Metabolic error
kinetics of enzyme is altered due to mutation
1. Abnormal response to activator fructose 1,6
bisphosphate
2. Abnormal Km or Vmax for substrate or coenzymes
3. Activity or stability of enzyme is altered
4. Amount of enzyme may be decreased
➢ PK deficiency is restricted to the Erythrocytes and
second most common cause of hemolytic anemia
after G-6 PD deficiency
 PYRUVATE KINASE DEFICIENCY AND HEMOLYTIC
ANEMIA (Cont)
PATHOGENESIS
➢ Mature erythrocytes are totally dependent on
glycolysis for ATP
➢ Na, K+ ATPase pump maintains biconcave shape of
R.B.C
➢ Without ATP-RBCs swell up and lyse,and can not
pass through capillaries and lyse
➢ Life span decreases and chronic hemolytic anemia
results
➢ Reticulocytes can survive but erythropoisis can not
cope with hemolysis
➢ Increased level of 2,3 BPG in R.B.C
 ALTERNATE FATES OF PYRUVATE

A. Oxidative decarboxylation of Pyruvate to Acetyl

CoA
➢ Major pathway in tissues with high oxidative capacity e.g
Cardiac Muscle
➢ Enzyme = Pyruvate dehydrogenas complex

B. Carboxylation of pyruvate to Oxaloacetate

➢ Provides intermediate for citric acid cycle
➢ Provides substrate for gluconeogenesis

C. Reduction of pyruvate to Ethanol (microorganism)

Metabolic fates of Pyruvate
 Step 11
REDUCTION OF PYRUVATE TO LACTATE

➢ Lactate is the final product of Anaerobic

glycolysis in eukaroytic cells

➢ Major fate in the R.B.C, WBC, Lens, Cornea,

Kidney medulla and Testes
 Reduction of pyruvate to lactate via
the enzyme lactate dehydrogenase.
(e.g., muscle during intense
activity) OR (retina, brain, RBCs).
NADH required for this reaction is supplied
by the 6th reaction (the dehydrogenation of
glyceraldehyde 3-phosphate).

Importantly, under anaerobic conditions,

the regeneration of NAD+ by this step is
essential for the continued functioning of
glycolysis.
 REDUCTION OF PYRUVATE TO LACTATE

A. Lactate Formation in muscle

(Severe exercise)
➢The oxidative capacity of the cell for NADH
produced by glycolysis and TCA is less due to
increased ratio of NADH / NAD+
➢More lactate is produced, pH is decreased and
muscle cramps
➢Lactate goes to liver and used by liver to make
glucose
How does fermentation
allow cells to produce
ATP in the absence of
oxygen?
Detoxifying of pyruvate
Regeneration of NAD

What happens to the

lactate?
Cori-cycle
Lactic Acidosis
 REDUCTION OF PYRUVATE TO LACTATE

B. Lactate Consumption (depends on NADH / NAD

reaction)
➢ In liver and Heart: The reaction of NADH / NAD is
lower than skeletal muscle
➢ Heart muscle oxidizes the lactate to Co2 and water
via Citric Acid Cycle
➢ Liver either oxidize to Co2 & H2o or convert it to
glucose via gluconeogenesis
 REDUCTION OF PYRUVATE TO LACTATE
C. Lactic Acidosis (Mainly due to Hypoxia / Anoxia of
tissues) Causes collapse of circulatory system
➢ Myocardial infartion
➢ Pulmonary embolism
➢ Uncontrolled haemorhhage
➢ Individual in shock

Pathogenesis
1. Inadequate oxygen supply to tissues leads to impaired
oxidative phosphorylation
2. Tissues survival dependent on Anaerobic glycolysis for ATP
with Lactic Acid as end product
3. Oxygen debt can be detected by blood Lactic Acid for
measuring the severely of shock and monitor the recovery
 ENERGY YIELD FROM GLYCOLYSIS
1. Anaerobic glycolysis
a. Only 2 – ATP
b. This is significant in following conditions
i. Limited oxygen supply
ii. Tissues with few or no mitochondria. e.g Kidney medulla,
mature Erythrocytes, Leukocytes and cells of Lens ,Cornea
and Testes
2. Aerobic Glycolysis
a. Consumed = 2 – ATP
b. Generated =
i. Substrate level = 4 ATP
ii. Oxidative Phosphorylation = (2 NADH) = 6 ATP
Total= 10 ATP
c. Net = 10 – 2 = 8 ATP
Energy calculation
 HORMONAL REGULATION OF GLYCOLYSIS

1. Short Term Regulation: (Minutes)

➢ Allosteric activation or inhibition and phosphorylation or
dephosphorylation of enzymes
2. Covalent modification, slower regulation of enzymes of
glycolysis & gluconeogenesis
3. Long Term Regulation:
[By induction via gene transcription] (Days)
➢ Regular Consumption of high carbohydrate meals causes
increased Insulin
➢ Insulin increases amount of enzymes in liver
▪ Glucokinase
▪ Phosphofructokinase
▪ Pyruvate kinase
➢ Glucagon decreases the gene transcription of these
enzymes
Hormonal regulation
 Conversion of Glucose to Fructose via Sorbitol

►Aldose reductase reduces glucose to sorbitol, which is

quite polar and thus does not passively diffuse across
membrane.
►Sorbitol is not a substrate for the glucose transporters.
►Therefore its trapped inside cells.
►Liver, ovaries, sperm, and seminal vesicles contain the
enzyme sorbitol dehydrogenase
►Oxidizes sorbitol to fructose.
►Fructose then enters glycolysis or gluconeogenesis.
 CLINICAL SIGNIFICANCE

►When glucose is elevated (e.g., diabetes) and if there is

sufficient NADPH, aldose reductase produces excess
sorbitol.
►Retina, lens, kidney, and nerve cells do not contain
sorbitol dehydrogenase and therefore sorbitol
accumulates.
►Causes a strong osmotic effect and cell swelling due to
water retention.
►Symptomalogy occurs (cataract formation, peripheral
neuropathy, and vascular problems).
No sorbitol
dehydrogenase.
Metabolic map
 CITRIC ACID CYCLE
(TCA CYCLE, KREBS CYCLE)
A process involving the oxidation of
Acetyl CoA to CO2 and H2O through a
series of reactions leading to the
production of ATPs
• O2 dependent
• Important intermediates
• Citrate: source for FA and Sterol synthesis
• α-KG and OA are sources for amino acids
• Succinyl CoA involved in porphyrin synthesis
 Tricarboxylic acid cycle
(Citric acid cycle or krebscycle)

• Plays several roles in metabolism

• Common metabolic pathway
• Occurs in mitochondrion and aerobic pathway
• Carbon skeleton of carbohydrates, amino acids
and fats are completely oxidized to water and
CO2
• Participates also in anabolic reactions
Oxidation decarboxylation of pyruvate to
form Acetyl CoA
1. Enzyme: Pyruvate dehydrognase complex
• Pyruvate is transported to mitochondria and
Is the main fuel for TCA or gluconeogenesis
a. Pyruvate dehydrogenase – (decarboxylase) = E1
b. Dihydrolipoyl transacetylase = E2
c. Dihydrolipoyl dehydrogenase = E3

2. Coenzymes
a. E1- requires – Thiamine pyrophosphate -TPP
b. E2- requires – lipoic acid and Coenzyme A
c. E3 requires FAD and NAD
Deficiency of Thiamine and Niacin cause serious central nervous
system problem
Steps in the conversion of Pyruvate to Acetyl CoA

1. Decarboxylation to form a hydroxyethyl derivative

bound to TPP

2. Intermediate is oxidized by lipoic acid

3. Transfer of acetyl group to CoA

4. Re-oxidation of lipoic acid (FAD dependent)

5. Reoxidation of FADH2 and reduction of NAD+ to form

NADH + H+
PYRUVATE DEHYDROGENASE COMPLEX
 Regulation of pyruvate dehydrogenase complex
E1: is regulated by cyclic AMP independent protein
kinase and phosphoprotein phosphatase. These are
part of enzyme complex

• Pyruvate inhibits it (product inhibition)

• Calcium is strong activator of phosphatase and
important in skeletal muscle
 Pyruvate Dehydrogenase deficiency

• Most common cause of Congenital lactic acidosis

• Main effect is on brain which relies for its ATP
from TCA
• This is biochemical basis of Beri Beri
• This may be most severe, moderate or mild
• This is x linked dominant disease
• Mild or expressed with episodic ataxia which is
induced by high carbohydrate diet
• Treatment by ketogenic diet, Low carbohydrate
and fat rich
 Arsenic poisoning of pyruvate Dehydrognase

• Thiol group of dihydrolipol transacetylase

• Arsenic also inhibits the glycolysis at glycaraldehyd-

3 phosphate

• Trivalent form of arsenite combines with thiol-SH

group of lipoic acid in enzyme E2

• Poisoning leads to neurological disturbances and

death
Succinate
thiokinase

CITRIC ACID CYCLE

 REACTIONS / STEPS OF CITRIC
ACID CYCLE
STEP - 1
1. Synthesis of citrate from acetyl CoA and
oxaloacetate

– Enzyme : citrate synthase

– Rate limiting and irreversible and regulated
step
– Regulated allosterically
– Activated by calcium and ADP
– Inhibited by ATP, NADH, succinyl CoA and
fatty acyl CoA
– Primarily by availability of substrates, acetyl
CoA and oxaloacetate
TCA CYCLE
Citrate
synthase
The citric acid cycle takes part in fatty
acid synthesis
 STEP 2

Citrate to cis-Aconitate
• Enzyme: Aconitase

• Reaction: Dehydration
Citrate is isomerized to isocitrate by this
first dehydration and yields cis-aconitate as
an intermediate.

• Prosthetic group: Fe-S

Aconitase
 STEP- 3
Formation of isocitrate from Cis -Aconitate
Enzyme = Aconitase

• Reversible step
• Inhibited / poisoned by fluoroacetate which is
converted to fluoroacetyl CoA, combines with
oxaloacetate to form fluorocitrate
• Flurocitrate is potent inhibitor of aconitase
Aconitase
 STEP - 4
Oxidation and decarboxylation of isocitrate to
form  - ketoglutarate

• Enzyme – isocitrate dehydrogenase

• Regulatory and irreversible step
• Yields one CO2 and NADH + H (3 ATP)
• Allosterically activated by ADP and Ca2+
• Inhibited by ATP and NADH (abundant energy
signal)
Isocitrate
Dehydrogenase
 STEP-5

Oxidative decarboxylation of  ketoglutarate to form

succinyl CoA
Enzyme  - ketoglutarate dehydrogenase complex

– Action similar to pyruvate dehydrogenase

– releases one – CO2 and one NADH (3ATP)
– Coenzymes = TPP, lipoic acid, FAD, NAD+, ad CO-A
– Irreversible and rate limiting step
– Enzyme – activated by – calcium
– Inhibited by ATP,GTP, NADH and succinyl co-A
– KG also comes from amino acids
α-ketoglutarate
dehydrogenase
 STEP- 6
Cleavage of succinyl CoA to form Sucinate
Enzyme = sucinate thiokinase

• Produces one GTP by substrate level

phosphorylation (ATP)
• Succinyl CoA is also produced from propionyl
CoA (odd number fatty acids) and amino acids
(valine, isoleuince, methionine)
Succinate
thiokinase
 STEP- 7
Oxidation of succinate to from fumarate

Enzyme = succinate dehydrogenase

Coenzyme = FAD and produces FADH
(2ATP)
Enzyme is inhibited by oxaloacetate
Succinate
dehydrogenase
 STEP- 8
Hydration of fumarate to form
malate

– Enzyme – Fumarase (Fumarate

hydratase)
– Freely reversible reaction
– Fumarate is also produced in urea cycle,
purine synthesis and from phenyl
alanine and tyrosine
Fumarase
 STEP- 9
Oxidation of malate to form
oxaloacetate

– Enzyme = malate dehydrogenase

– Coenzyme NAD- produces NADH + H (3
ATP)
– Oxaloacetate is also produced from
aspartate by transamination
Malate
Dehydrogenase
 Salient features of citric acid cycle
• Provides substrate for the respiratory chain in
the form of NADH + H and FADH + H
• Produces two molecules of CO2 per cycle
• One oxaloacetate enters the cycle and
regenerated at the end
• Furnace for burning of acetyl – CoA
• Vitamins play a key role in the citric acid cycle

– Thiamine, Niacin (NAD), Riboflavin

(FAD),Pantothenic acid (CoA)
Twelve ATP are formed per turn of the citric acid cycle
 ENERGY BALANCE SHEET
• Aerobic Glycolysis NADH + H+ produced = 1 x 2
ATPs produced = 2 x 2
ATPs utilized = 2 x 1
Total = 8 ATPs
• Citric Acid Cycle NADH + H+ produced = 3 x 2
FADH2 produced = 1 x 2
ATPs produced = 1 x 2
Total = 12 x 2 = 24
• Pyruvate to Acetyl CoA
NADH + H+ produced = 1 x 2 = 3
3x2=6
• Total ATPs produced by complete oxidation of 1 molecule of
glucose:
Malate shuttle: 38 ATPs
Glycerophosphate shuttle: 36 ATPs
 Regulation of the citric acid cycle

A. Regulation by activation and inhibition of three

enzymes

1. Citrate synthase
2. Isocitrate dehydrogenase
3. -ketoglutarate dehydrogenase
 GLUCONEOGENESIS
• Some tissues depend completely on glucose
eg., brain, nervous system, RBCs, testes, renal
medulla and embryonic tissue.
• Brain: 120 g of glucose/day (more than ½
stored glucose).
• Glycogen depleted: between meals, fasts,
vigorous exercise etc.
 GLUCONEOGENESIS
Definition
Synthesis of glucose from non carbohydrate sources
▪ Liver glycogen Can provide the glucose only for 10-
18 hours
▪ After 18 hours gluconeogenesis is the source of
glucose
▪ Brain, erythrocytes, kidney medulla, lens, cornea,
testes and exercising muscle are totally dependent
on glucose as fuel
▪ Liver, kidney and enterocytes are the main sites of
gluconeogenesis
▪ During overnight fast 90% of glucose is formed by
liver and 10% from kidney during prolonged fast
▪ 40% glucose from kidney afterwards
 Substrates for Gluconeogensis

• Glycerol, lactate, and the keto acids

obtained from transamination of
glucogenic amino acids

A. Glycerol
– From hydrolysis of triacylglycerol and
conversion to dihydroxyacetone phosphate
in liver which joins glycolysis
– Adipocytes lack glycerol kinase and can not
use glycerol
 SYNTHESIS OF GLUCOSE FROM GLYCEROL
• Glycerol released from hydrolysis of TAGs on adipose
tissues
• Transported from adipose tissues to liver

Glycerol kinase
(a) Glycerol Glycerol phosphate
Glycerol phosphate
(b) Glycerol phosphate dehyrogenase
Dihyroxyacetone phosphate

Note: Adipocytes DO NOT have glycerol kinase

and thus cannot phosphorylate glycerol
SYNTHESIS OF GLUCOSE FROM LACTATE
B. Lactate

➢ Produced in the exercising skeletal muscle and is

taken up by liver and converted into glucose and
added to blood
➢ Also produced in tissues without mitochondrion,
mainly erythrocytes
C. Amino Acid
➢ Is major source during fasting which give rise to
Pyruvate, succinyl CoA, Oxaloacetate, α-
ketoglutarate and fumarate by transamination of
glucogeneic amino acids

D.Propionate, end product of odd number fatty

acids
 Relation between Glycolysis and Gluconeogenesis
– Not identical but run in opposite directions.
– 7 out of 10 reactions of glycolysis are reversible
– 3 irreversible steps:

Hexokinase
1. Glucose Glucose 6-Phosphate

PFK1
1. Fructose 6-Phosphate Fructose 1,6-
bisphosphate

Pyruvate kinase
1. Phosphoenolpyruvate Pyruvate
 Reactions unique to gluconeogenesis

• Seven glycolytic reactions are reversible

and are used in the synthesis of glucose
from lactate and Pyruvate

• Three irreversible reaction are

circumvented by four alternate reactions
SYNTHESIS OF GLUCOSE FROM PYRUVATE

Pyruvate is converted to phosphoenol pyruvate

in 2 exergonic reactions:

Pyruvate
Carboxylase
(a) Pyruvate Oxaloacetate

(b) Oxaloacetate PEP CK Phosphoenolpyruvate

Pyruvate carboxylase utilizes Biotin as prosthetic group.
Biotin carboxylation is catalyzed at one active site of
Pyruvate Carboxylase.

Biotin + ATP + HCO3- → carboxybiotin + ADP + Pi

At the other active site of Pyruvate Carboxylase, the

activated CO2 is transferred from biotin to pyruvate

Carboxybiotin + pyruvate biotin + oxaloacetate

A. Carboxylation of Pyruvate to Oxaloacetate in
mitochondria

➢ First road block is to overcome the irreversible

reaction of Pyruvate to Phosphoenol Pyruvate (PEP)
➢ Occurs in mitochondria of liver and kidney

1. Enzyme: Pyruvate Carboxylase (coenzyme is Biotin)

➢ This reaction provides oxaloacetate for
gluconeogenesis
2. Pyruvate Carboxylase is allosterically regulated
➢ Activated by increased acetyl CoA
➢ Inactivated by decreased level of acetyl CoA
Acetyl CoA
+
allosterically
regulates
pyruvate
carboxylase
 Transport of Oxaloacetate to cytosol

➢Further enzymes of gluconeogenesis are

present in cytosol

➢It is transported in the form of malate to

cytosol and converted back to oxaloacetate

➢α-Ketoglutarate, succinyl CoA, fumarate,

intermediates of TCA derived from amino
acids are also converted to oxaloacetate
B. Decarboxylation of Cytosolic oxaloacetate to
form Phosphoenol Pyruvate (PEP)

➢ Enzyme = Phosphoenol pyruvate carboxykinase

(PEPCK)
➢ 1 mole of GTP is used in this reaction
➢ PEP is then converted to fructose 1,6
bisphosphate by reversible reactions of
glycolysis
 C. Dephosphorylation of fructose 1,6 bisphosphate to
form Fructose – 6 - Phosphate
Enzyme = Fructose 1,6 bisphosphatase
➢ This is important regulatory site of gluconeogenesis and
glycolysis
1. Regulation by energy levels with in the cell
➢ Energy poor state i.e increased AMP inhibit it
➢ Energy rich state (high ATP) increases its activity
➢ β-Oxidation is mandatory for running gluconeogenesis
2. Regulation by Fructose 2,6 Bisphosphate
Inhibits this enzyme and concentration of Fructose 2,6
Bisphosphate is decreased by Glucagon during fasting
3. Fructose 6-Phosphate is converted to glucose 6-Phosphate
by isomerase by reversible reaction
Dephosphorylation of Fructose 1, 6 Bisphosphate
Effects of elevated level of Glucagon
 Dephosphorylation of Glucose
6-Phosphate to form glucose

Enzyme = Glucose 6-Phosphate Phosphatase

Site: Liver , Kidney, can only release free glucose

into the blood
➢ Muscle cells also lack this enzyme and can not
produce glucose
➢ Deficiency of this enzyme results into von
gierk’s disease
3. Dephosphorylation of Glucose 6-phosphate to
Glucose:
(a) Glucose 6-phosphate enters the ER by

Glucose 6-phosphate translocase

Glucose 6-
(b) Glucose 6-phosphate phosphatase Glucose
Dephosphorylation of Glucose 6 Phoasphate
 Summary of the reactions of glycolysis and
gluconeogenesis

Enzymes of Irreversible Circumvented by

Reactions
➢ Hexokinase / glucokinase
➢ Phosphofructokinase (PFK)
➢ Pyruvate Kinase
➢ Glucose 6-Phosphatase
➢ Fructose 1,6-
Bisphosphatase
➢ Pyruvate carboxylase
PEP Carboxy kinase
ENERGY USED IN GLUCONEOGENESIS

Six high energy phosphate bonds

and oxidation of 2 x NADH are
required for the formation of each
molecule of glucose
ENERGY USED IN GLUCONEOGENESIS

Six high energy phosphate bonds

and oxidation of 2 x NADH are
required for the formation of each
molecule of glucose
GLUCONEOGENESIS
REGULATION OF GLUCONEOGENESIS
Regulation depends upon

1. Circulating level of glucagon (Hormone)

2. Availability of substrates of
gluconeogenesis

3. Altered rate of synthesis and degradation

affect enzyme activity
 Regulation of Gluconeogenesis
• Hormonal Glucagon activates PEP CK
Glucagon inhibits Hexokinase,
PFK-2 & Pyruvate kinase

• Allosteric activation: Acetyl CoA (Pyruvate

carboxylase)

• Allosteric inhibition AMP (Fructose 1,6

Bisphosphatase)
A. Glucagon
1.Changes in Allosteric effectors of
enzymes
1.Glucagon lowers the level of fructose
2,6-bisphosphate resulting in activation
of F 1,6- bisphosphate and inhibition of
PFK-1
2.Glucagon increases protein kinase via
cyclic AMP and inactivates pyruvate
kinase
3.Increases the transcription of PEP
carboxykinase gene by induction
Regulation by Glucagon
 Allosteric inhibition by AMP

➢ AMP – inhibits Fructose 1,6-Bisphosphatase and

activates phosphofructokinase (PFK-1)

➢ Elevated AMP level stimulates the oxidation of

Acetyl CoA by β-oxidation and produces ATP and
NADH required for gluconeogenesis
 PENTOSE PHOSPHATE PATHWAY
(HEXOSE MONOPHOSPHATE, 6-PHOSPHOGLUCONATE
PATHWAY)
• Occurs in the cytosol
• No ATP formed or utilized
• 2 NADPH produced per glucose 6-phosphate
• Also produces ribose 5-phosphate (nucleotide
biosynthesis)
• Consists of 2 major portions
• 2 irreversible oxidative reactions
• Series of reversible non-oxidative reactions
 TK TA
TK

G6PD 6PGD
 NADPH
• Phosphate group confers specificity and selectivity
for enzymes
• Functions of NADPH
• Reductive Biosynthesis
• Reduction of Hydrogen Peroxide
• Cytochrome P450 Monooxygenase
system
• Phagocytosis by WBCs
• Synthesis of NO
 REDUCTIVE BIOSYNTHESIS
• A high energy molecule involved in
metabolic processes by virtue of its ability
to act as a reducing agent.
• Synthesis of
• Fats
• Steroids
• cholesterol
 ROLE WITH ANTIOXIDANTS

Reactive Oxygen Species and specific antioxidant enzymes

GLUTATHIONE

NADPH and
GLUTATHIONE
 CYTOCHROME P450 MONOOXYGENASE
SYSTEM

• Consists of mixed function oxidases.

• Adds 1 atom of O2 to substrate and the other
to water (reduced in the presence of NADPH).
R-H + O2 + NADPH + H+
R-OH + H2O + NADP+
Comprises of two systems (depending on site)
• Mitochondrial system
• Hydroxylation of steroids.
• Synthesis of steroid hormones.
• Synthesis of bile acid.
• Synthesis of 1,25 hydroxycholecalciferol.
• Microsomal system
• Found in SER of liver.
• Detoxifies drugs, pesticides, carcinogens etc.
 PHAGOCYTOSIS BY WBCS

• Receptor mediated endocytosis of foreign

particles by cells like monocytes.
• Oxygen dependent (myeloperoxidase
system).
• Oxygen independent system.
SYNTHESIS OF NITRIC OXIDE
 GLUCOSE 6-PHOSPHATE DEHYROGENASE
DEFICIENCY
• X-linked enzyme deficiency disorder.
• Most common in Middle East and Africa.
• Defect in the enzyme leads to defect in the cell’s
ability to detoxify oxidizing agents.
• Leading to cell death and hemolysis (RBCs).
• Precipitated by drugs (sulfa drugs, primaquin and
acetanilid).
• Resistance against falciparum malaria in female
carriers.
GLUCOSE 6-PHOSPHATE METABOLISM IN RBCS
 FRUCTOSE METABOLISM
• Fructose is a ketohexose.
• Mainly found in fruits and other sources
of sucrose.
• Absorption and entry into cells is
independent of insulin.
• Requires GLUT 5 transporters.
 Hereditary Fructose Intolerance
• Seen in 1:20,000 live births.
• Deficiency of aldolase B.
• At the time of weaning (exposure to fructose and
sucrose).
• Fructose 1-phosphate accumulates.
• ATP level falls ----hypoglycemia and vomiting.
• Decreased protein synthesis ---- bleeding etc.
• Liver failure and death.
• Diagnosis: Fructosuria and RFLP test.
 GALACTOSE METABOLISM
• An aldohexose obtained from lactose.

• First undergoes phosphophorylation.

• Conversion to UDP-galactose (uridine

diphosphate-galactose).
• UDP-galactose used in biosynthetic reactions.

• Converted to UDP-glucose for glycolysis and

gluconeogenesis.
 Classic Galactosemia

• Autosomal recessive disorder

• Deficiency of Galactose 1-phosphate uridyl transferase
• Accumulation of galactose 1-phosphate and galactose
• Similar picture as in HFI but more tissues affected
• Severe mental retardation, liver damage, cataract.
• Diagnosis: antenatal chorionic villus sampling,
galactosuria
• Treatment: removal of galactose and lactose from the
diet.
 LACTOSE METABOLISM
• Galactosyl β (1 4) glucose
• Synthesis:
• In the endoplasmic reticulum
• Lactose synthase (UDP-galactose: glucose
galactosyltransferase
• Enzyme has 2 proteins
• Protein A (β-D-galactosyltransferase) to GlcNAC
• Protein B (α lactalbumin) mammary gland glucose
• Lactose Intolerance
 GLYCOGEN METABOLISM
• Storage form of glucose.
• Branched chain polymer of α–D-
Glucose.
• Liver glycogen maintains blood glucose
level.
• Muscle glycogen fuel reserve for muscle
contraction.
 GLYCOGENESIS
• The synthesis of glycogen from glucose.
• Glycogenesis occurs in the cytosol in the presence
of ATP and UTP.
• Steps of glycogen synthesis:
• Synthesis of UDP glucose.
• Synthesis of a primer (Glycogenin)
• Chain elongation
• Branch formation
 Synthesis Of UDP-Glucose
1. Conversion of glucose 1-phosphate to glucose 6-
phosphate (phosphoglucomutase).
2. Synthesis of UDP-glucose from glucose 1-
phosphate and UTP (UDP-glucose
pyrophosphorylase).
Glucose 6-phosphate

UTP + Glucose 1-phosphate

UDP-Glucose + PPi
 Synthesis of a primer (Glycogenin)

• A protein compound.

• The OH-group of tyrosine accepts the 1st

glycosyl unit.

• Self promoting attachment of first few units.

• Stays associated to the molecule and found in

the centre
 Chain elongation
• Transfer of glucose from UDP-glucose.
• By glycogen synthase.
• Α (1 4) linkage is created.

Branch formation

Transfer of 5-8 glycosyl residues after every 8

residues
Amylo-α (1 4) α (1 6) transglucosidase
 GLYCOGENOLYSIS
• The process of breakdown of glycogen to glucose.

• Takes place in the cytosol by a separate set of

enzymes.

• Chain shortening.

• Removal of branches.

• Conversion of glucose 1-phosphate to glucose

6-phosphate.
 Chain shortening
• Removal of glucose 1-phosphate from
glycogen.

• Enzyme: Glycogen phosphorylase.

• Coenzyme: Pyridoxal phosphate.

• Site of attack: α(1 4) linkage

• Continues till 4 glycosyl units remain.

• Resulting compound: Limit dextran.

Cleavage of an alpha (1 4) glycosidic bond
 Removal of branches
• Carried out by debranching enzyme.
• It has 2 domains
• Oligo α(1 4) α(1 4)-glucan transferase
• For removal of first three glycosyl residues.
• Amylo α(1 6)-glucosidase
• For removal of last glycosyl residue
• The straight chain is acted upon by glycogen
phosphorylase.
 Conversion of glucose 1-phosphate

to glucose 6-phosphate

• Conversion by phosphoglucomutase

• Glucose 6-phosphate enters the ER by glucose 6-

phosphate translocase

• Glucose released by glucose 6-phosphatase

 Regulation of Glycogen synthesis
• Liver glycogensis starts after food intake whereas,
glygogenolysis takes place in fasting.
• Muscle glygogenolysis starts after exercising
whereas glycogensis takes place in resting skeletal
muscle.
• Regulation:
• Allosteric regulation
• Glycogen synthase and glycogen
phosphorylase
• Hormonal regulation
• Glycogen synthase
Allosteric
Regulation of
Glycogen
Synthesis and
Breakdown
DEGRADATION OF GLYCOGEN
IN MUSCLES BY CALCIUM

Phosphorylase
kinase
Hormonal activation of Glycogen degradation
GLYCOGEN STORAGE DISEASES

A group of inherited disorders resulting

from defects in enzymes required either
for the normal synthesis or breakdown of
glycogen characterized by deposition of
abnormal type or quantity of glycogen.
 GLYCOGEN STORAGE DISEASE (GSD)
1. Von Gierke’s Disease – Type-I
Type = I-a = Glucose 6-phosphatase deficiency
Type = II-b = Glucose 6-phosphate translocase deficiency
➢ Affects Liver, Kidney and Intestine
➢ Fasting hypoglycemia is severe
➢ Fatty liver, hepatomegaly
➢ Progressive renal damage
➢ Growth retardation and delayed puberty
➢ Hyper lacticacidemia
➢ Hyper lipidemia
➢ Microscopically normal glycogen structure and increased
glycogen stores
➢ Treatment
Nocturnal gastric infusions of glucose
2. Pompe Disease Type-II

Enzyme = Lysosomal α (1-4) Glycosidase deficiency

➢ Inborn lysosomal enzyme defect

➢ Generalized (Primarily structure liver, heart,
muscle)
➢ Excessive normal glycogen found in abnormal
vacuols in cytosol
➢ Normal blood sugar levels
➢ Massive cardiomegaly & death occurs due to heart
failure
3. Cori’s disease or Limit dexpinosis = Type-III
➢ Enzyme = Debranching enzyme
➢ Site = Liver and muscle
➢ Glycogen phosphorylase is present and acts on glycogen
to form limit dextrin
➢ Limit dextrin accumulates in liver and muscle – shows
mild Type-I symptoms

4. Anderson’s disease or Amylopectinosis = Type-IV

➢ Enzyme = Branching enzyme
➢ Site = Liver
➢ Abnormal glycogen deposited in liver
➢ Disease presents by cirrhosis , leading to death
5. MC Ardle’s disease Type -V
Enzyme = Skeletal muscle Phosphorylase deficiency
➢ Liver enzyme normal
➢ Temporary weakness and cramping of skeletal muscles
after exercise
➢ Normal mental development
➢ No rise in blood lactate during strenuous exercise
➢ Myoglobinemia and myoglobinuria
➢ High level of glycogen with normal structure in muscle

6. HER’s disease Type - VI

➢ Enzyme deficient = liver phosphorylase
➢ Hypoglycemia
GLYCOGENOSIS NAME CAUSE OF
DISORDER
Type I Von Gierke’s Deficiency of G6Pase

Type II Pompe’s Defidiency of lysosomal

acid maltase
Type III Forbes’, Cori’s Absence of debranching
enzyme
Type IV Amylopectinosis Absence of branching
enxyme
Type V McArdle’s Absence of muscle
phosphorylase
Type VI Hers’ Deficiency of liver
phosphorylase
Type VII Tauri’s Deficiency of PFK in
liver and RBCs
 URONIC ACID PATHWAY
• Oxidation of glucose at different C-atoms leads to the
production of different acid sugars.
• Uronic acid pathway in the liver converts glucose to:
• Glucuronic acid
• Ascorbic acid
• Pentoses
• Importance:
• Precursors of proteoglycans
• Precursors of conjugated glucuronides (steroids,
bilirubin and drugs)
 Active compound

Gulonolactone oxidase
 SIGNIFICANCE OF URONIC ACID PATHWAY
• Major pathway for the synthesis of
glucuronides.
• Excretion of toxic metabolites.
• Chemicals (xenobiotics).
• Other wastes.

• Enzyme deficiency can lead to Essential

pentosuria leading to excessive excretion of
L-xylulose in urine.
 GAGs
►Animal heteroglycans/Mucopolysaccharides
►Unbranched—linear chains of repeating
disaccharide units
– Amino sugar D-glucosamine or D- galactosamine
– Uronic acid L glucuronic acid or L iduronic acid
except keratan sulphate
Except hyaluronic acid all GAGs contain S gp
►GAG is a polymer of [amino sugar+uronic A]n
GAGs in Proteoglycan aggregate
 GAGs
►Core protein-
– extracellular,
– covalent,
– proteoglycan except hyaluronic acid
►Bottle brush
►A molecule of hyaluronic acid to form aggregate
►Ionic bond b/w core protein and hyaluronic acid
►This association further supported by link proteins
►Amount of CHO in glycoproteins is upto 95%
SYNTHESIS OF AMINO SUGARS
SYNTHESIS OF ACIDIC SUGARS
 Occurrence-GAGs

►Synovial fluids of joints

►Vitreous humour of eye
►Arterial walls
►ECF
►Bones
►Cartilages
 GAGs- functions
►Major component of EC matrix
►Sulphate & COO gps impart neg charge and
ability to hold water→gel→cushion against
mechanical shock
►Molecular sieves→ control entry and exit of
substances from cells
►Elasticity
►Lubricate joints
►Viscous lubricating properties of mucous
secretions →thus name mucopolysaccharides
 GAGs types

►Different types based upon

– Uronic acid comp
– Amino sugar comp
– Linkage b/w amino and uronic acid
– Chain length of di- sacch polymer
– Presence or absence of Sulphate group and its
position of attachment to sugar
– Nature of core protein
– Tissue, subcellular distribution, role
Chondroitin Sulphate
Hyaluronic Acid
Heparin
Name Disaccharide unit Unique features/ function

hondroitin sulfate N acetyl galactosamine Most prevalent

Glucuronic acid GAG,endoskeleton, elasticity of
cartilage

Dermatan sulfate N acetyl galactosamine Transparency of cornea

L iduronic acid

eratan sulfate N acetyl glucosamine Transparency of cornea

Galactose ( no uronic acid)

Heparin Glucosamine Anticoagulant, Highest negative

Glucuronic/iduronic acid charge density of any known
biological molecule

Heparan sulfate Same as heparin some Receptor, cell growth, charge

glucosamine are acetylated selectivity of glomerular
membrane
 Glycoconjugates
► Proteoglycans
– much larger CHO chains which are negatively
charged

►Glycoproteins
– mainly proteins,
– <4% CHO att to polypeptide chain as oligo chains
which are mostly
►Branching
►Don’t contain repeating disaccharide units
►May or may not negatively charged
CHO in Glycoproteins
 Glycoproteins
– Glycans covalently attached to polypeptide
chains
– Attachment as O- glycosidic to side chain O of
serine & threonine
– Att as N glucosidic to side chain N of asparagine
– L fucose and N acetylglucosamine appearing at
ends
– called glycosylation- post translational
modification

► Mucoproteins >4% CHO

– Found in joints for lubrication
 Functions- Glycoproteins
►Almost all plasma proteins except albumin
►Many integral membrane proteins
►Most of secreted proteins
– Antibodies
– Hormones
►Clotting factors
►Serve as lubricant and protective agents
– Gastric mucin
– Urogenital secretions
►Transport molecules
– Transferrin
– Ceruloplasmin
 Cont..

►CHO chains with these proteins serve as/to

– Regulate lifespan of proteins

( removal of terminal sialic acid residue-
removal of RBC from circulation)
– Recognition signal for cell-cell interaction
( sperm-oocyte)
– Stabilize proteins against denaturation
– Facilitate solubility of proteins
Maj M Javad Yousaf Asst Prof AM
264
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 GLUCOSE HOMEOSTASIS

• Maintenance of normal blood glucose level is called Glucose

Homeostasis
• Carbohydrates are the main source of energy in the body &
glucose is the most preferred source of energy for most of the
body tissues,
• Brain tissues derive the energy mainly from glycogen.

• Normal fasting plasma glucose level is (60-100mg%)

• Normal random plasma glucose level is (130-150mg%)

Maj M Javad Yousaf Asst Prof AM

265
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• Glucose Homeostasis in the body is
maintained by two sets of factors, some
factors add while other remove glucose
from the blood & balance of these two
factors keeps the blood glucose level at a
fairly constant level.

Maj M Javad Yousaf Asst Prof AM

266
College
Addition Factors in Blood
• Absorption from GUT
• Hepatic Glycogenolysis
• Gluconeogenesis
• Renal Reabsorption
• Glucose obtained from other sources

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267
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 Removal Factors in Blood
• Hepatic Glycogenesis
• Muscle Glycogenesis
• Lipogenesis
Minor Pathways for Glucose oxidation:
– HMP shunt
– Uronic Acid pathway
Major Pathway for Glucose oxidation:
– Glycolysis
– Citric Acid Cycle
Synthesis of amino acids:
from intermediates of glycolysis

268
M
 Dietary Carbohydrate

Glucose Glycogen
Fructose
Galactose
Amino acids
Glycerol
Lactate
e sis s
en ysi
og n ol
yc e
Gluconeogenesis Gl c og
y
Gl
NADPH

HMP
Carbon skeleton Glucose Pathway
Ribose-P
of amino acids

Gluconeogenesis
Energy

Glycolysis
Other
Carbohydrates Glycerol-P

Pyruvate
Fatty Acids

ATP Acetyl-CoA
ADP+Pi
Triacylglycerol
Electron
Transport
Chain
NADH Kreb's Cycle

Maj M Javad Yousaf Asst Prof AM

2CO2 269
College
Maj M Javad Yousaf Asst Prof AM
270
College