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
