Glycolysis
Definition: Glycolysis means oxidation of glucose to give
pyruvate (in the present of oxygen (aerobic)
, or lactate in the absence of oxygen (anerobic).
The reactions of Glycolysis take place in the cytosol of all
tissue cells.
It is of physiological important in:
1-Tissue with no mitochondria: mature RBCs, Cornea and lens
(anerobic).
2-Tissues with few mitochondria: Testis, leucocytes, medulla
of the kidney, retina, skin and gastrointestinal tract.
3-Tissue undergoes frequent oxygen lack: skeletal muscle
especially during exercise.
Under anaerobic conditions:
Glycolysis consists of 11 coupled reactions with the
overall net reaction being:
D-glucose + 2 Pi + 2 ADP ------------→2 lactate + 2 H+ + 2
ATP + 2 H2O
Under aerobic conditions:
Glycolysis consists of 10 coupled reactions with the
overall net reaction being:
D-glucose + 2 Pi + 2 ADP + 2 NAD+--------→2 pyruvate + 2
ATP + 2 NADH H+
Glycolytic Pathway
Glycolysis represents the central pathway for the catabolism of
glucose.
Glycolysis is a catabolic pathway occurs in the cytoplasm
that is found in all cells —aerobically or anaerobically.
Glycolysis involves ten reactions - same in all cells, but rates
differ- (eleven reactions in anaerobic condition)
Two phases:
First phase converts glucose to two Glyceraldehyde-3-phosphate
Second phase produces two pyruvate
Aerobically; products are pyruvate, ATP and NADH
Anaerobicaly; the products are lactate and ATP
Glucose enters the Glycolysis pathway by conversion to
glucose-6-phosphate. Initially, there is energy input
corresponding to cleavage of two ~P bonds of ATP.
1-Hexokinase
catalyzes:
glucose + ATP glucose-6-phosphate + ADP
I- HEXOKINASE - GLUCOKINASE
Catalyze 1st Reaction in glycolytic pathway:
Glucose enters cell and is phosphorylated to glucose 6-
phosphate
Reaction is irreversible.
Hexokinase active at low glucose to meet needs of brain
cells and RBCs
Glucokinase in liver must handle the increase in glucose
after a meal
Glucokinase, present in liver and β-cells
COMPARING HEXOKINASE & GLUCOKINASE
Hexokinase (and glucokinase) act to phosphorylate glucose
into glucose-6-P and keep it in the cell
Hexokinase (Km glucose = 0.1 mM); cell has 4 mM glucose.
So hexokinase is normally active!
Hexokinase is regulated - allostercally inhibited by (product)
glucose-6-P
Hexokinase can also phosphorylate fructose, mannose,
and glucosamine, whereas glucokinase cannot. It has high
Km for fructose,
Glucokinase (Km glucose = 10 mM) only turns on when cell is
rich in glucose- feeding state Glucokinase is activated by
insulin in liver & pancrease
Km Michaelis constant : amount of substrate that give half
of maximum velocity
Hexokinas and Glucokinase
Glucokinase HEXOKINASE
Liver and β-cells Brain cells and RBCs Site
Activated with insulin Normally active Activity
-Ve +Ve Fructose
Phosphorylation
Active at high glucose Active at low glucose Glucose level
level level
2. Phosphoglucose Isomerase
Catalyzes:
Glucose-6-phosphate fructose-6-phosphate
(aldose) (ketose)
3. Phosphofructokinase
catalyzes:
fructose-6-phosphate + ATP fructose-1,6-bisphosphate +ADP
This highly spontaneous reaction has a mechanism similar to
that of Hexokinase.
The Phosphofructokinase reaction is the rate-limiting step of
Glycolysis. The enzyme is highly regulated.
Phosphofructokinase (PFK; PFK1)
PFK-1 is the major point of regulation in glycolysis. It is
irreversible. PFK is regulated allosterically.
Glucagon inhibit phosphofructokinase PFK-1
4. Aldolase
catalyzes:
fructose-1,6-bisphosphate dihydroxyacetone
phosphate + glyceraldehyde-3-phosphate.
The Aldolase reaction is an aldol cleavage, the reverse
of an aldol condensation
Phase one
5. Triosephosphate Isomerase (TIM)
catalyzes:
Dihydroxyacetone phosphate (ketose)
glyceraldehyde-3-phosphate (aldose)
6. Glyceraldehyde-3-phosphate Dehydrogenase
catalyzes:
glyceraldehyde-3-phosphate + NAD+ + Pi
1,3,bisphosphoglycerate + NADH + H+
7. Phosphoglycerate Kinase
1,3-bisphosphoglycerate + ADP
3-phosphoglycerate + ATP
8. Phosphoglycerate Mutase
catalyzes:
3-phosphoglycerate 2-phosphoglycerate
Phosphate is shifted from the hydroxyl on C3 of 3-
phosphoglycerate to the hydroxyl on C2
9. Enolase
catalyzes:
2-phosphoglycerate phosphoenolpyruvate +
H2O
This Mg++-dependent dehydration reaction is inhibited by
fluoride. Fluorophosphate forms a complex with Mg++ at
the active site.
10. Pyruvate Kinase
catalyzes:
phosphoenolpyruvate + ADP pyruvate + ATP
This transfer of phosphate from PEP to ADP is
spontaneous. yields an unstable enol, that
spontaneously converts to the keto form of pyruvate.
Required inorganic cations K+ and Mg++ bind to anionic
residues at the active site of Pyruvate Kinase.
Pyruvate Kinase
Transfer of a phosphoryl group from
phosphoenolpyruvic to ADP catalyzed by pyruvate
kinase.
Irreversible; An important site of regulation in the liver.
Site of synthesis of ATP“substrate level
phosphorylation”,
Pyruvate kinase is activated by ; insulin and Fructose
1,6 Bi-Posphate
and inhibited by; ATP
Phase two
REGULATION OF GLYCOLYSIS
Glucagon ; decrease in PFK1 activity, nactivate
pyruvate kinase thereby decreasing glycolysis.
Insulin causes increasing glycolysis by
A .Activation of pyruvate kinase
b. Activation of glucokinase
ATP inhibits both phosphofructo-kinase and pyruvate
kinase, so, excess ATP production inhibits further
glycolysis.
glucose + 2 NAD+ + 2 ADP + 2 Pi
2 pyruvate + 2 NADH + 2 ATP
Three enzymes catalyze highly spontaneous reactions:
Key enzymes of glycolysis
Hexokinase,
Phosphofructokinase, (rate limiting)
Pyruvate Kinase.
Control of these enzymes determines the rate of the
Glycolysis pathway.
Local control involves dependence of enzyme-catalyzed
reactions on concentrations of pathway substrates or
intermediates within a cell.
Global control involves hormone-activated production of
second messengers that regulate cellular reactions for the
benefit of the organism as a whole.
