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CARBOHYDRATE

METABOLISM
CATABOLISM
EDITED BY
Liniyanti D.Oswari,MD.,MNS,MSc.
For Block 8
Medical student, Sriwijaya
University

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

lactate

acetyl CoA

EtOH

Glucose
Utilization
Adipose

Energy
Stores

Glycogen

Glucose
Pentose
Phosphate
Pathway

Ribose-5-phosphate

Glycolysis

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-6phosphate (G-6-P).
Other monosaccharides are also made to G6-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).

Hexokinase
ADP

Glucose

ATP

Fructose-6-phosphate
ATP

ADP

Fructose-1, 6-biphosphate
Dihydroxyacetone
phosphate (DHAP)

Glyceraldehyde-3phosphate
NAD + Pi
NADH + H+

ADP

Glyceraldehyde-1,
3-bisphosphate
ADP

Glycerol ADP

ATP

ATP

H2O Phospho

ATP

Glycerate-3phosphate

Pyruvate

Glucose-6phosphate

Glycogen
Lactate
Dehydrogenase

Glucose-1phosphate

UDP-glucose

Lactate

Glycolysis:
break down of glucose in cytoplasm

Glycerate-2phosphate

-enolpyruvate
H2 O

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 Cs) is
split into two 3-carbon compounds:
Glyceraldehyde 3-phosphate (GAP)

Glycolysis: Phase 3
Phase 3: Oxidation and ATP formation
The 3-carbon sugars are oxidized (reducing
NAD+); i.e., 2 Hs + NAD
NADH2
Inorganic phosphate 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 payof phase.
Reactions 6-10. two glyceraldehyde
3-phosphate molecules to two
pyruvate plus four ATP molecules.
-- A net of two ATP molecules overal
plus 2 NADH(10 ATP2 ATP= 8 ATP).

GLYCOLYSIS

Glucose
ATP
hexokinase
ADP
Glucose 6-phosphate
phosphoglucoisomerase
Fructose 6-phosphate
ATP
phosphofructokinase
ADP
Fructose 1.6-bisphosphate
aldolase
triose phosphate isomerase
Dihydroxyacetone
Glyceraldehyde

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

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
Low Vmax

High Km
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.

GLUCOKINASE

liver enzyme with high KM (10 mM)fo


glucose so most effective when
glucose levels are very high
not inhibited by glucose 6-phospha
sensitive to high glucose in
circulation from recent meal
so it decreases high level of glucose
in blood by taking glucose into liver

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 v
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 1,6bisphosphate


- (feed-forward)
Phosphorylation (inactive form) and
dephosphorylation (active form)
under hormone control.

Glycolysis:
Embden-Myerhof
Oxidation of
Pathway
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 1,6
bisphosphate

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

Net = 2 ATP and 2 NADH( 8 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
energy from glucose.

Carbohydrate Metabolism

Primarily glucose

All cells can utilize glucose for energy production

Fructose and galactose enter the pathways at various points


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

Fates of Glucose

Fed state

Storage as glycogen

Storage as lipids

Liver
Skeletal muscle
Adipose tissue

Fasted state

Metabolized for energy


New glucose synthesized

Synthesis
Synthesis and
and
breakdown
breakdown occur
occur
at
at all
all times
times
regardless
regardless of
of
state...
state...
The
The relative
relative rates
rates
of
of synthesis
synthesis and
and
breakdown
breakdown change
change

High Blood Glucose

Pancreas
Muscle
Glucose
absorbed

Insulin

Glycogen

Glucose
absorbed

Adipose
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

2nd Priority: provide energy

Stored in muscle and liver


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)

Fate of Product of GlycolysisPyruvate


- 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.

Cori Cycle
Lactate is
converted
to pyruvate
in the liver

Two anerobic pathways:

- to lactate via lactate dehydrogenase


- to ethanol via ethanol dehydrogena
- 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
C

CH 3

COO
Alanineaminotransferase
(AAT)

Pyruvate

Keto acid
acid

HC

NH 3+

CH 3
Alanine

Amino

Pyruvate Dehydrogenase Complex


(PDH)

Prepares pyruvate to enter the TCA cycle

Aerobic Conditions
Electron
Transport

TCA
Cycle

1. Lactate Fermentation
Enzyme = Lactate
Dehydrogenase
COOC=O + NADH + H+
NAD+
CH3

pyruvate
lactate

COOH-C-OH +
CH3

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

-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-P
glycogen
glycog

ATP
ATP
NADH
Blood
NAD
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
COOC=O
CH3

CO2

CH2OH
H O

C + NADH
CH3 +
CH3
NAD+

pyruvate
acetaldehyde
ethanol
pyruvate decarboxylase-

- 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 aceta
- 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
anaerobic
2 ethanol + CO2

2 NADH
anaerobic
2 lactate

2 acetyl CoA + 2
CO2

O2

aerobic

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.

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

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 - Fig. 23.18

a phosphoanhydride exchange

driven by pyrophosphate hydrolysis

Glycogen Synthase
Forms alfa-(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 Ser14 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

HormonalGlucagon
Regulation
II
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

CH2OH
O

CH2OH

CH2
O

O
O

-[1-6] linkage
CH2OH
O
O

-[1- 4] linkages

. The glycogen structure showing the glycosidic bonds

Glycogenesis

Liver

710% 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
Hexokinase
(muscle)
Glucokinase
(liver)

ATP
ADP

Glucose-6-phosphate

Phosphoglucomutase

(Glucose)

(Glucose)n+

Glucose-1-P
Uridyltransferase

UDP-glucose

Glucose-1-phosphate
UTP

UDP

Glycogen Synthase

PPi

Pathway of glycogen synthesis (glycogenesis).

Control of enzyme activity

Rate limiting step

Glycogen synthesis

Glucose 6-P glucose 1-P.


glucose 1-P + UTPUDP-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

Protein phosphatase 1 (PP1) regulates


glycogen metabolism.

Inactive

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
Glucose-1-phosphate
phosphoglucomutase

LIVER PATHWAY
glucose-6-phosphatase

Glucose-6-phosphate
glycolysis
(inhibited by lack of
fructose-2,6-bisP

Glycogenolysis and the fate of glycogen in liver and kidney

Glucose
Pi

Glycogen

MUSCLE PATHWAY
Pi

glycogen
phosphorylase
Glucose-1-phosphate
phosphoglucomutase
Glucose-6-phosphate
glycolysis
Pyruvate
pyruvate
dehydrogenase
Acetyl CoA

anaerobic
Lactate
lactate dehydrogenase
citric acid cycle
aerobic

CO2

. 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 and glucagon stimulate
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


Glucose6phosphat
e

6Phosphogluconolactone

6Phosphogluconate
D-Ribulose5phosphate

RNA or
DNA

D-Ribose5phosphate

Oxidative branch

ATP

ADP

Glucose-6-P-dehydrogenase
NADP
NADPH
6-Phosphogluconate

Glucose 6-P

Glucose

NADP

6-Pgluconate dehydrogenase

Glyceraldehyde 3-P

TDP
Fructose 6-P

Transketolase

Non-oxidative branch

Ribulose 5-P

Xylulose 5-P
Glyceraldehyde 3-P

CO2

NADPH

Ribose 5-P (5 carbons)


Transketolase
TDP
Sedoheptulose 7-P (7 carbons)
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

Glyceraldehyde 3-P

TDP
Fructose 6-P

Transketolase

Ribulose 5-P

Xylulose 5-P
Glyceraldehyde 3-P

Ribose 5-P (5 carbons)


Transketolase
TDP
Sedoheptulose 7-P (7 carbons)
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

ATP
Glucose

ADP

Glucose-6-P-dehydrogenase
NADP
NADPH
6-Phosphogluconate

Glucose 6-P
Ribulose 5-P

NADP
6-Pgluconate dehydrogenase

CO2

NADPH

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

Allosteric Regulation

First step
Rate limiting
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
Cells 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

isomerization stage

generation of NADPH
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

6-phosphoglucono is hydrolyzed (addition of water) to 6phosphogluconate


6-phosphogluconate is oxidized by 6 phosphogluconate
dehydrogenase

produces NADPH and 6-phosphoglucono

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

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

PFK, PDH, CS, and isocitrate dehydrogenase

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