BIOC405 Fall 2013

Monday October 28th

 Last time:
Principles of Metabolism
ATP, NADH, Creatine Phosphate
Coupled Reactions

Introduction to Glycolysis

Today:
10 steps to Glycolysis
Fate of pyruvate

What is Glycolysis?
Uses Glucose:
Corn syrup, sucrose, starch, glycogen
Only source of energy for the brain
Fuels quick bursts of activity in muscle

10 reactions
Takes place in the cytoplasm of a cell.
The oxidation of glucose to pyruvate, NADH
and ATP.

Overall reaction of glycolysis:

Glucose + 2 Pi + 2 ADP + 2 NAD+

2 pyruvates + 2 ATP + 2 NADH + 2 H+ + 2 H2O

Phosphoglucose isomerase 2nd step in

Glycolysis
aldose

ketose

Phosphoglucose Isomerase Reaction 2

Glucose-6-P to Fructose-6-P
Why does this reaction occur?
Next step (phosphorylation at C-1) would be tough for
hemiacetal OH but easy for primary OH
Isomerisation activtes C-3 for cleavage in aldolase reaction.

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Phosphofructokinase 3rd step in Glycolysis

ATP
Phosphofructokinase I
ADP
PFK-I and PFK-II play a
central role in
glycolysis regulation
(more on this later).
Note difference between bis phosphate vs. di phosphate
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12

Aldolase 4th step in Glycolysis

3
1

2
4

3
5

2
3

Splits bond between carbons 3 and 4.

13

Aldolase and Triose

Phosphate
Isomerase
4th and 5th steps in
Glycolysis
Aldolase

Step 5

Triose phosphate isomerase (TIM)

14

The
Investment
Phase

Glucose
ATP
hexokinase
phosphoglucose isomerase
ATP

phosphofructokinase
aldolase

Dihydroxyacetone
Phosphate

Glyceraldehyde 3 P
TIM
ATP

Driving force for the TIM reaction is

the siphoning off of Glyceraldehyde
3 P into the final stages of glycolysis.

ATP
Pyruvate

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GAPDH and phosphoglycerate

kinase
6h and 7th steps in
Glycolysis
Glycolysis
NAD+
G = -1.1 kJ/mol

Pi

Glyceraldehyde 3-phosphate NADH

+H+
dehydrogenase
GAPDH

Phosphoglycerate kinase
PGK

ADP
ATP

This type of ATP production is called

SUBSTRATE-LEVEL PHOSPHORYLATION
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The GAPDH and PGK reactions are coupled

to drive production of both NADH and ATP.

GAPDH

GAP + Pi + NAD+

1,3-Bisphosphoglycerate + NADH Go = +6.7 kJ/mol

1,3-BPG + ADP

3-Phosphoglycerate + ATP Go= -18.8kJ/mol

PGK

-----------------------------------------------------------------------------------------GAP +Pi + NAD+ + ADP

3PG + NADH +ATP

G0= -12.1kJ/mol

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Phosphoglycerate mutase8th step in Glycolysis

Phosphoglycerate mutase (PGM)

19

mutases cause
intramolecular shift
of a chemical group

The process involves a

2,3-bisphosphate intermediate.

Where have you seen this molecule before?????

20

8 mM
4 mM

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Box 7-3

Hyperventilation raises pH of blood.

Respiratory Alkalosis
To the air
lungs
H2O +

CO2

H2CO3

H+ + HCO3-

Bicarbonate
buffer in
blood serum

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Synthesis of 2,3 bisphosphoglycerate in ERYTRHOCYTES

The amount of 2,3
BPG in erythrocytes
increases when the
pH of blood
increases

Bisphosphoglycerate mutase is
primarily found in erythrocytes.
Box 15-2 figure 1

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Inherited diseases involving 2,3 BPG

Less than normal 2,3 BPG)

More than normal 2,3 BPG)

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Box 15-2 figure 2

Enolase and Pyruvate

Kinase9th and 10th
steps in Glycolysis
Enolase

Pyruvate kinase

H2O

ADP

ATP
Substrate level phosphorylation again

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26

Pyruvate kinase (phosphoryl transfer).

-16 kJ/mol

-46 kJ/mol

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Stryer 5/e

The
Investment
Phase

Glucose
ATP
hexokinase
phosphoglucose isomerase
ATP

phosphofructokinase
aldolase

Dihydroxyacetone
Phosphate

Glyceraldehyde 3 P
TIM
ATP

Driving force for the TIM reaction is

the siphoning off of Glyceraldehyde
3 P into the final stages of glycolysis.

ATP
Pyruvate

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Glucose
ATP
ATP
DHAP
2 NADH
The
Payoff
Phase

2 ATP

Glyceraldehyde 3 P
GAPDH
PGK
PGM
Enolase

2 ATP

Pyruvate Kinase
Pyruvate

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A balanced equation for Glycolysis

Glucose + 2 Pi + 2 ADP + 2 NAD+


2 pyruvate + 2 ATP + 2 NADH + 2 H+ + 2 H2O
Problem: How to regenerate NAD+?

30

NAD+
needed
here:

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Anaerobic fate of
pyruvate (muscle)

This reaction is referred to as homolactic fermentation

Glycolysis to lactate is referred to as anaerobic glycolysis
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Anaerobic fate of
pyruvate (yeast)

TPP

This reaction is referred to as alcoholic fermentation

33

Structure & Function of Thiamine Pyrophosphate.

The cofactor, TPP is used as a coenzyme for decarboxylations of keto acids. It is derived from thiamine (Vitamin B1) by transfer of a
pyrophosphate group from ATP to thiamine, yielding TPP and AMP.

We also will encounter TPP in pyruvate dehydrogenase (PDH) and as

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an aldehyde carrier in transketolase reactions.

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Anaerobic fate of
pyruvate (yeast)

TPP

This reaction is referred to as alcoholic fermentation

Aldehyde dehydrogenase
Alcohol dehydrogenase
ethanol
acetaldehyde
CH3CH2OH + NAD+
CH3CHO + H+ + NADH
acetaldehyde
acetate
CH3CHO + NAD+ + H2O
CH3COO- + NADH + 2H+
Aldehyde dehydrogenase
2 forms of aldehyde
dehydrogenase mitochondrial
and cytoplasmic
Some people (particularly Asians) have a less active form of
the mitochondrial form of aldehyde dehydrogenase.
Acetaldehyde accumulates in blood.

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Disulfiram: inhibitor of
acetaldehyde dehydrogenase
Discovered by accident in 1948
while looking for a remedy for
parasite infections. Investigators
tried it on themselves and found
they could not drink alcohol.!
!
Now used as a treatment for
alcoholism.!

Flushing of face!
Headache!
Nausea!
Chest pain!
Vomiting!
Blurred vision!
Mental confusion!
etc....!
!

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Some Key Concepts for Glycolysis

Sources of energy, ATP, Phosphocreatine, glycolysis,
respiration!
Types of Glucose Transporters and hexokinases!
Importance of coupled reactions!
Overall scheme for glycolysis!
10 steps of glycolysis and their significance!
Investment vs. Payoff phases!
Isomerase vs. Mutase!
Significance of 2,3 BPG!
Which steps use and which steps make ATP!
How NADH is regenerated in animals vs. in yeast.!
How alcohol is made and how it is metabolized.!
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