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at The End of This Lecture, Student Will Be Able To
at The End of This Lecture, Student Will Be Able To
at The End of This Lecture, Student Will Be Able To
8
Glycolysis
1
Faculty of pharmacy ©M. S. Ramaiah University of Applied Sciences
Glycolysis
2
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
6 CH OPO 2
2 3
5 O
H H
H
4 H 1
OH
OH OH
3 2
H OH
glucose-6-phosphate
3
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
Types of glycolysis
4
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
6 CH 2 O H 6 CH O PO 2
2 3
ATP ADP
5 O 5 O
H H H H
H H
4 1 4 H 1
OH H OH
M g 2+
OH OH OH OH
3 2 3 2
H OH H ex ok inase H OH
glucose glucose -6 -ph osp hate
1. Hexokinase catalyzes:
Glucose + ATP glucose-6-P + ADP
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++.
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Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
NH2
ATP N
N
adenosine triphosphate
O O O N N
O P O P O P O CH2 adenine
O
H H
O O O
H H
OH OH
ribose
6
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
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 OH
Mg2+
OH OH OH OH
3 2 3 2
H OH Hexokinase H OH
glucose glucose-6-phosphate
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Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
6 CH 2 O H 6 CH O PO 2
2 3
ATP ADP
5 O 5 O
Induced fit: H H H H
H H
4 1 4 H 1
OH H 2+ OH
Glucose binding OH OH
Mg
OH OH
3 2
to Hexokinase 3 2
H ex o k in ase
H OH H OH
stabilizes a glu co se glu cose -6 -ph osp hate
conformation
in which: glucose
Hexokinase
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Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
6 CH O PO 2
2 3
6 CH 2
5 O 2 O PO 3 1 CH 2 O H
H H O
H
4 H 1 5 H HO 2
OH
OH OH H 4 3 OH
3 2
OH H
H OH
P h o s p h o g lu c o s e Is o m e ra s e
g lu c o s e -6 -p h o s p h a te fru c to s e -6 -p h o s p h a te
H 4 3 OH M g2+ H 4 3 OH
OH H OH H
fru c to s e -6 -p h o s p h a te f r u c t o s e - 1 ,6 - b i s p h o s p h a t e
3. Phosphofructokinase catalyzes:
fructose-6-P + ATP fructose-1,6-bisP + 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, as will be discussed later
Faculty of Pharmacy
11
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
2
1 CH 2 O PO 3
2C O
H O
2
HO 3 C H A ld o la se 3
CH 2 O PO 3 1C
H 4C OH 2C O + H 2C O H
2
H C OH 1 CH 2 O H 3 CH 2 O PO 3
5
2 d ih y d ro x y a c e to n e g ly c e ra ld e h y d e -3 -
6 CH 2 O PO 3
p h o sp h a te p h o sp h a te
fru c to se -1 ,6 -
b isp h o sp h a te
T rio se p h o sp h a te Iso m e ra se
Faculty of Pharmacy
12
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
lysine 1CH2OPO3
2
H
2C NH (CH2)4 Enzyme
H 3N +
C COO +
HO 3
CH
CH2
H 4
C OH
CH2
H 5
C OH
CH2 2
6 CH 2OPO 3
CH2
Schiff base intermediate of
NH3 Aldolase reaction
2C O
H O
2
HO 3 C H A ld o la se 3
CH 2 O PO 3 1C
H 4C OH 2C O + H 2C O H
2
H C OH 1 CH 2 O H 3 CH 2 O PO 3
5
2 d ih y d ro x y a c e to n e g ly c e ra ld e h y d e -3 -
6 CH 2 O PO 3
p h o sp h a te p h o sp h a te
fru c to se -1 ,6 -
b isp h o sp h a te
T rio se p h o sp h a te Iso m e ra se
d ih y d ro x y a c e to n e e n e d io l g ly c e ra ld e h y d e -
p h o s p h a te in te rm e d ia te 3 -p h o s p h a te
g ly c e ra ld e h y d e - 1 ,3 -b is p h o s p h o -
3 -p h o s p h a te g ly c e ra te
6. Glyceraldehyde-3-phosphate Dehydrogenase
catalyzes:
glyceraldehyde-3-P + NAD+ + Pi
1,3-bisphosphoglycerate + NADH + H+
Faculty of Pharmacy
17
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
G ly c e ra ld e h y d e -3 -p h o s p h a te
D e h y d ro g e n a se
H O + H+ O O PO 3 2
1 C NAD+ NADH 1 C
+ Pi
H C OH H C OH
2 2
2 2
3 CH 2 O PO 3 3 CH 2 O PO 3
g ly c e ra ld e h y d e - 1 ,3 -b is p h o s p h o -
3 -p h o s p h a te g ly c e ra te
HC CH CH 2OPO 3 2
glyceraldehyde-3-
OH OH phosphate
Oxidation to a Enz-Cys S CH CH CH 2OPO 3 2
carboxylic acid NAD + thiohemiacetal
(in a ~ thioester) NADH intermediate
occurs, as NAD+ O OH
is reduced to Enz-Cys S C CH CH 2OPO 3 2
NADH. Pi
acyl-thioester
intermediate
O OH
2
Enz-Cys SH O 3PO C CH CH 2OPO 3 2
1,3-bisphosphoglycerate
The “high energy” acyl thioester is attacked by Pi to
yield the acyl phosphate (~P) product. 20
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
H O O
H H
C C
NH2 NH2
+
N + N
2e + H
R R
NAD+ NADH
Faculty of Pharmacy
21
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
P h o s p h o g ly c e ra te K in a s e
2
O O PO 3 ADP ATP O O
1C 1
C
H 2
C OH H 2
C OH
2+
2 Mg 2
3 CH 2 O PO 3 3 CH 2 O PO 3
1 ,3 - b i s p h o s p h o - 3 -p h o s p h o g ly c e ra te
g ly c e ra te
Faculty of Pharmacy
23
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
P h o s p h o g ly c e r a te M u ta s e
O O
histidine
O O
H
1
C 1
C
2 H3N+ C COO
H 2C O H H 2 C O PO 3
2 CH2
3 CH 2 O PO 3 3 CH 2 O H
C
3 -p h o s p h o g l y c e r a t e 2 -p h o s p h o g ly c e r a te
HN CH
HC NH
An active site histidine side-chain
participates in Pi transfer, by
O O
donating & accepting phosphate. C
1
The process involves a H 2C OPO32
2,3-bisphosphate intermediate. 3CH2OPO3
2
2,3-bisphosphoglycerate
View an animation of the
Phosphoglycerate Mutase reaction.
Faculty of Pharmacy
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Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
E n o la s e
O H
O OH O
O O O
C C 1
C
1
H 2 C O P O 32 C O P O 32 2C O P O 32
3 C H 2O H C H 2O H 3CH2
2 -p h o s p h o g ly c e ra te e n o la te in te r m e d ia te p h o s p h o e n o lp y r u v a te
9. Enolase catalyzes:
2-phosphoglycerate phosphoenolpyruvate + H2O
This dehydration reaction is Mg++-dependent.
2 Mg++ ions interact with oxygen atoms of the substrate
carboxyl group at the active site.
The Mg++ ions help to stabilize the enolate anion
intermediate that forms when a Lys extracts H+ from C #2.
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Faculty
Faculty of Pharmacy
of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
P y r u v a te K in a s e
O O O O
ADP ATP
1
C 1
C
2
C O PO 3 2 2
C O
3 CH 2 3 CH 3
p h o s p h o e n o lp y r u v a te p y r u v a te
Faculty of Pharmacy
26
Faculty of Pharmacy ©M. S.©M. S. Ramaiah
Ramaiah University
University of Applied
of Applied Sciences
Sciences
P y r u v a te K in a s e
O O O O O O
C ADP ATP C C
1 1 1
2
2
C O PO 3 2
C OH 2
C O
3 CH 2 3 CH 2 3 CH 3
p h o s p h o e n o lp y ru v a te e n o lp y ru v a te p y ru v a te
glyceraldehyde-3-phosphate + dihydroxyacetone-phosphate
Triosephosphate
Isomerase
Glycolysis continued
Faculty of Pharmacy
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©M. S. Ramaiah University of Applied Sciences
glyceraldehyde-3-phosphate
NAD+ + Pi Glyceraldehyde-3-phosphate
NADH + H+ Dehydrogenase
1,3-bisphosphoglycerate
Glycolysis ADP
continued. Phosphoglycerate Kinase
ATP
Recall that 3-phosphoglycerate
there are 2 Phosphoglycerate Mutase
GAP per 2-phosphoglycerate
glucose.
H2O Enolase
phosphoenolpyruvate
ADP
Pyruvate Kinase
ATP
pyruvate 29
Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences
Summary
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Faculty of Pharmacy ©M. S. Ramaiah University of Applied Sciences