MUHAMMAD WAJID

Institute of Pharmacy Gulab Devi Educational

Complex, Lahore
 Metabolism:
2

 Metabolism is “the entire set of enzyme- catalyzed

transformations of organic molecules in living cells”

 Two broad classes

 Catabolism & Anabolism Catabolic Pathways

 Transform fuels into cellular energy

Muhammad Wajid
 Conti…
3

 Requires inputs of energy to proceed

 Useful energy + small molecules complex molecules

 Pathways that can be either anabolic or catabolic,

depending on the energy conditions in the cell are
referred to as amphibolic pathways

Muhammad Wajid
 Glycolysis:
4

 Derived from Greek word Glykys (Sweet) and Lysis

(Splitting)
 The process in the cell metabolism by which
carbohydrates and sugar, especially glucose, are
broken down, producing ATP and pyruvic acid and two
“high energy” electron carrying molecules of NADH
 During this process one molecule of glucose (6 carbon
molecule) is degraded into two molecules of pyruvate
(3 carbon molecule)
 It is also called as Embden-Mayerhof Parnas Pathway
or EMP pathway
Muhammad Wajid
 Conti…
5

 It was the first metabolic sequence to be studied

 It take place in all body cells

 It occurs in cytosol

 The enzymes of this pathway are present in the

cytosomal fraction of the cell

Muhammad Wajid
 Introduction:
6

 Glycolysis is the sequence of 10 enzyme-catalyzed

reactions that converts glucose into pyruvate with
simultaneous production of ATP
 In this oxidative process, 1molecule of glucose is
partially oxidized to 2 molecules of pyruvate
 This major pathway of glucose metabolism occurs in
the cytosol of all cell
 This unique pathway occurs aerobically as well as
anaerobically & doesn’t involve molecular oxygen
Muhammad Wajid
 Conti…
7

 The glycolytic sequence of reactions differ from

species to species only in the mechanism of its
regulation & in the subsequent metabolic fate of the
pyruvate formed

 In aerobic organisms, glycolysis is the prelude to Citric

acid cycle and ETC

 Glycolysis is the central pathway for Glucose

catabolism
Muhammad Wajid
 Conti…
8

 Glycolysis occurs in the absence of oxygen

(Anaerobic) or in the presence of oxygen (Aerobic)

 Lactate is the end product under anaerobic

conditions

 In aerobic condition, pyruvate is formed, which is

then oxidized to CO2 and H2O

Muhammad Wajid
 Two phases of glycolysis:
9

 Glycolysis leads to breakdown of 6-C glucose into

two molecules of 3-C pyruvate with the enzyme
catalyzed reactions being bifurcated or categorized
into 2 phases:

o Phase 1- preparatory phase

o Phase 2- payoff phase

Muhammad Wajid
 Preparatory phase:
10

 It consists of the 1st 5 steps of glycolysis in which the

glucose is enzymatically phosphorylated by ATP to
yield Fructose-1,6-bisphosphate

 This fructuse-1,6-biphosphate is then split in half to

yield 2 molecules of 3-carbon containing
Glyceraldehyde-3-phosphate/ dihyroxyacteone
phosphate

Muhammad Wajid
 Conti…
11

 Thus the first phase results in cleavage of the

hexose chain

 This cleavage requires an investment of 2 ATP

molecules to activate the glucose molecule and
prepare it for its cleavage into 3-carbon
compound

Muhammad Wajid
 Payoff phase:
12

 This phase constitutes the last 5 reactions of Glycolysis

 This phase marks the release of ATP molecules during

conversion of Glyceraldehyde-3-phosphtae to 2
molecules of Pyruvate

 Here 4 moles of ADP are phosphorylated to ATP.

Although 4 moles of ATP are formed, the net result is
only 2 moles of ATP per mole of Glucose oxidized,
since 2 moles of ATP are utilized in Phase 1

Muhammad Wajid
 Stepwise explanation of glycolysis:
13

 Phosphorylation:
 Glucose is phosphorylated by ATP to form glucose-
6- phosphate
 This is an irreversible reaction & is catalyzed by
hexokinase
6 CH2OH 6 CH OPO 2
2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H 2+ OH
Mg
OH OH OH OH
3 2 3 2
H OH Hexokinase H OH
glucose glucose-6-phosphate
Muhammad Wajid
 Hexokinase:
14

 The reaction involves nucleophilic attack of the C6

hydroxyl O of glucose on P of the terminal phosphate
of ATP

 ATP binds to the enzyme as a complex with Mg++

 Mg++ interacts with negatively charged phosphate

oxygen atoms, providing charge compensation &
promoting a favorable conformation of ATP at the
active site of the Hexokinase enzyme

Muhammad Wajid
 Isomerization:
15

 It is a reversible rearrangement of chemical

structure of carbonyl oxygen from C1 to C2,
forming a Ketose from the Aldose
 Thus, isomerization of the aldose Glucose 6-
phosphate gives the ketose, Fructose-6-phoshphate
6 CH OPO 2
2 3
5 6 CH OPO 2 1 CH2OH
H O H 2 3
O
H
4 H 1 5 H HO 2
OH
OH OH H 4 3 OH
3 2
OH H
H OH
Phosphoglucose Isomerase
glucose-6-phosphate fructose-6-phosphate
Muhammad Wajid
 Phosphorylation:
16

 Here the Fructose-6-phosphate is phosphorylated by

ATP to fructose-1,6-bisphosphate
 This is an irreversible reaction and is catalyzed by
phosphofructokinase enzyme
Phosphofructokinase
6 CH OPO 2 1CH2OH 6 CH OPO 2 1CH2OPO32
2 3 2 3
O ATP ADP O
5 H HO 2 5 H HO 2

H 4 3 OH Mg2+ H 4 3 OH
OH H OH H
fructose-6-phosphate fructose-1,6-bisphosphate
Muhammad Wajid
 Breakdown:
17

 This six carbon sugar is cleaved to produce two 3-

C molecules: glyceradldehyde-3-phosphate (GAP)
& dihydroxyacetone phosphate(DHAP)
 This reaction is catalyzed by Aldolase
2
1 CH2OPO 3

2C O
H O
2
HO 3 C H Aldolase 3
CH2OPO3 1C

H 4C OH 2C O + H 2
C OH
2
H C OH 1CH2OH 3 CH2OPO 3
5
2
6 CH2OPO 3 dihydroxyacetone glyceraldehyde-3-
phosphate phosphate
fructose-1,6-
bisphosphate
Triosephosphate Isomerase
Muhammad Wajid
 Isomerization:
18

 Dihydroxyacetone phosphate is oxidized to form

Glyceraldehyde-3-phosphate
 This reaction is catalyzed by triose phosphate
isomerase enzyme
Triosephosphate Isomerase
H H OH H
+
O
+ + +
H C OH H H C H H C
C O C OH H C OH
CH2OPO32 CH2OPO32 CH2OPO32

dihydroxyacetone enediol glyceraldehyde-

phosphate intermediate 3-phosphate
Muhammad Wajid
 Phosphorylation:
19

 This is the first reaction of energy yielding step.

 Oxidation of aldehyde derives the formation of a
high energy acyl phosphate derivate

Glyceraldehyde-3-phosphate
Dehydrogenase
H O + H+ O OPO 32
1C NAD+ NADH 1 C
+ Pi
H C OH H C OH
2 2
2 2
3 CH2OPO 3 3 CH2OPO 3

glyceraldehyde- 1,3-bisphospho-
3-phosphate glycerate

Muhammad Wajid
 Conti….
20

 An inorganic phosphate is incorporated in this

reaction without any expense of ATP
 NAD+ is the cofactor in this reaction which acts as an
oxidizing agent
 The free energy released in the oxidation reaction is
used in the formation of acyl phosphate

Muhammad Wajid
 Dephosphorylation:
21

 This reaction generates ATP from the energy

released by the oxidation of 3-Phosphoglycerate
 The enzyme undergoes substrate-induced
conformational change similar to that of Hexokinase
Phosphoglycerate Kinase
O OPO 32 ADP ATP O O
1C 1
C
H 2
C OH H 2
C OH
2+
2 Mg 2
3 CH2OPO 3 3 CH2OPO 3

1,3-bisphospho- 3-phosphoglycerate
glycerate
Muhammad Wajid
 Phosphoglycerate Mutase:
22

 Phosphate is shifted from the OH on C3 to the OH on

C2

Phosphoglycerate Mutase
O O O O
C
1
C
1
H 2C OH H 2C OPO32
2
3 CH2OPO3 3 CH2OH
3-phosphoglycerate 2-phosphoglycerate
Muhammad Wajid
 Enolase Reaction:
23

Enolase
O  H  O  OH O
O O O
C C 1
C
1
H 2 C OPO32 C OPO32 2C OPO32

3 CH2OH CH2OH 3 CH2

2-phosphoglycerate enolate intermediate phosphoenolpyruvate

Muhammad Wajid
 Pyruvate Kinase:
24

Pyruvate Kinase
O O O O
ADP ATP
1
C 1
C

2
C OPO32 2
C O

3 CH2 3 CH3
phosphoenolpyruvate pyruvate

Muhammad Wajid
 Conti…
25

glucose Glycolysis
ATP
Hexokinase
ADP
glucose-6-phosphate
Phosphoglucose Isomerase
fructose-6-phosphate
ATP
Phosphofructokinase
ADP
fructose-1,6-bisphosphate
Aldolase

glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
Muhammad Wajid
 Conti….
26

glyceraldehyde-3-phosphate
NAD+ + Pi Glyceraldehyde-3-phosphate
NADH + H + Dehydrogenase
1,3-bisphosphoglycerate
ADP
Phosphoglycerate Kinase
ATP
3-phosphoglycerate
Phosphoglycerate Mutase
2-phosphoglycerate

H2O Enolase
phosphoenolpyruvate
ADP
Pyruvate Kinase
ATP
pyruvate
Muhammad Wajid
 Regulation of glycolysis:
27

 The rate of glycolysis is regulated by controlling of the 3

irreversible enzymes (key enzymes)
 These enzymes catalyze what is called committed reactions of the
pathway
 These enzymes are
o glucokinase (hexokinase)
o phospho-fructokinase-1
o pyruvate kinase

Muhammad Wajid
 Hormonal regulation:
28

 Insulin: Stimulates synthesis of all key enzymes of

glycolysis
 It is secreted after meal (in response to high blood
glucose level)
 Glycogen:
 Inhibits the activity of all key enzymes of glycolysis.
 It is secreted in response to low blood glucose level
Muhammad Wajid
 Energy regulation:
29

 High level of ATP inhibits PFK-1 and pyruvate kinase

 High level of ADP and AMP stimulate PFK-1

Muhammad Wajid
 Substrate regulation:
30

 Glucose-6-phosphate inhibits hexokinase (and not

glucokinase)

 Fructose 2,6 bisphosphate stimulates

phosphofructokinase-1

 Citrate inhibits phosphofructokinase-1

 Fructose 1,6 bisphosphate stimulates pyruvate kinase

Muhammad Wajid
31

Muhammad Wajid