GLYCOLYSIS

Catabolism of Glucose

Important Infos regarding Glycolysis

It happens in the cytoplasm (cytosol) It will occur even without oxygen Glucose = raw material (fuel) Consists of 2 phases: 1st phase = energy investment phase; 2nd phase = energy production phase Glyceraldehyde-3-phosphate (G3P) is the product of the 1st phase. One mole of glucose yields 2 moles of G3P. In the 1st phase, 2 ATP are consumed. In the 2nd phase, 4 ATP are produced for a net yield of 2 ATP for the whole process Main product of the whole process in aerobic environment (ample oxygen) = 2 Pyruvate (pyruvic acid) Main product in anaerobic environment (lack or inadequate O2) = 2 Lactic acid (lactate) Pyruvic acid (pyruvate) can enter the mitochondria and there, it is oxidized to Acetyl CoA The NADH produced in the cytosol by glycolysis (and perhaps by other reactions also) can also enter the mitochondria (albeit indirectly) using either the glycerol phosphate shuttle mechanism or the malate-aspartate shuttle. If the NADH uses the glycerol-phosphate shuttle, it has to give its hydrogen to FAD of the ETC. But if it uses the malate-asparate shuttle, it will enter the ETC via the NAD gate.

STRUCTURAL PROPERTIES OF PHOSPHATE

Phosphate is phosphoric acid dissolved in water. Phosphoric acid has 3 acidic hydrogens (triprotic). All the 3 detach from the structure when dissolved in water, the remaining structure is the phosphate ion. When acidic hydrogen detaches, the portion of the structure where it cleaves becomes full negatively charged. Thus, the oxidation number of phosphate is 3. Phosphate is a resonance structure

Structure of Phosphoric acid and phosphate

O
H

P O
H

Phosphoric acid Phosphate ion

1ST PHASE: ENERGY INVESTMENT PHASE

Final Product of the 1st phase

2nd Phase: Energy Production phase

Final product

Reactions of Glycolysis
1. Phosphorylation of glucose *hexokinase *glucokinase 2. Isomerization of glucose-6phosphate *phosphoglucose isomerase 3. Phosphorylation of fructose-6phosphate *phosphofructokinase (PFK1) 4. Cleavage of fructose 1,6biphosphate *Aldolase A 5. Isomerization of dihydroxyacetone phosphate *triose phosphate isomerase 6. Oxidation of glyceraldehydes 3phosphate *glyceraldehydes 3-phosphate dehydronase 7. Formation of ATP from 1,3biphosphoglycerate and ADP *phosphoglycerate kinase 8. Shift of the phosphate group from C3 to C2 *Phosphoglycerate mutase 9. Dehydration of 2phosphoglycerate *enolase 10. Formation of Pyruvate *pyruvate kinase 11. (Reduction of pyruvate to lactate) *lactate dehydrogenase

Mechanism of
Glycolysis

H C H C C C C C H

PHOSPHORYLATION OF GLUCOSE
Phosphorylation = attaching phosphate group to a chemical structure ATP-dependent (needs ATP for the rxn to occur) Enzymes: Hexokinase and Glucokinase Irreversible

OH H OH OH

HO H H H

P OH

GLUCOSE 6PHOSPHATE (G6P)

H H C C C C C C H OH

H HO H H H

O OH H OH OH

ISOMERIZATION OF GLUCOSE-6GLUCOSEPHOSPHATE
These 2 groups exchange position

Enzymes: Hexokinase and Glucokinase Reversible

FRUCTOSE-6-PHOSPHATE GLUCOSEGLUCOSE

H H C C C C C C H

OH P

O H OH OH

PHOSPHORYLATION OF FRUCTOSE-6FRUCTOSEPHOSPHATE
Phosphorylation = attaching phosphate group to a chemical structure ATP-dependent (needs ATP for the rxn to occur) Enzymes: Phosphofructokinase-1 (PFK-1) Rate-limiting enzyme Irreversible

HO H H H

FRUCTOSE 6PHOSPHATE 6 FRUCTOSE1,6BIPHOSPHATE

H H C C HO C

OH P

CLEAVAGE OF FRUCTOSE-1,6FRUCTOSE-1,6BIPHOSPHATE

H H H

C H C C H

DIHYDROXYACETONE PHOSPHATE (DHAP) H Fructose-1,6-biphosphate breaks into 2. H C == O OH OH Readily reversible Glyceraldehyde-3C OH H OH phosphate is the raw material for the 2nd phase C P P H of glycolysis Enzyme = aldolase H GLYCERALDEHYDE3PHOSPHATE (G3P) GLYCERALDEHYDE

FRUCTOSE1, 6BIPHOSPHATE FRUCTOSE 6

H H C == O OH C --------OHO C H H H H C == O OH C C H OH

ISOMERIZATION OF DHAP
DIHYDROXYACETONE PHOSPHATE (DHAP) Reversible DHAP is transformed to G3P Enzyme = triose phosphate isomerase Isomerase = class of enzyme that transfers group from one portion to another portion of the same substance GLYCERALDEHYDE GLYCERALDEHYDE 3PHOSPHATE (G3P)

H------

C=O is oxidized to carboxyl. Then the OH of carboxy is replaced by phosphate group

OXIDATION OF G3P
Reversible Main product = 1,3BPG Other products = NADH Enzyme = Glyceraldehyde-3phosphate dehydrogenase

H O C

P O OH

H H

C C H

OH

1,31,3GLYCERALDEHYDE GLYCERALDEHYDE 3BIPHOSPHOGLYCERATE PHOSPHATE (G3P) (1,3(1,3-BPG)

Phosphate cleaves and replaced by OH. The acidic hydrogen of OH dissociates, leaving O-

O C

O P-H

FORMATION OF ATP FROM 1,3-BPG

Reversible Main product = 3phosphoglycerate (3-PG) Other products = ATP (from phosphorylation of ADP) Enzyme = Phosphoglycerate kinase Kinase = enzymes used to attach or detach phosphate group

H H

C C H

OH
P

1,31,33-PHOSPHOGLYCERATE BIPHOSPHOGLYCERATE (3(3 (1,3- -PG) (1,3-BPG)

The two groups exchange position

O C

O-

SHIFT OF PHOSPHATE GROUP FROM C3 TO C2

H H

C C H

P OH OH P

Reversible Main product = 2phospholgycerate Enzyme = Phosphoglycerate mutase (an isomerase)

2-PHOSPHOGLYCERATE (23 PHOSPHGLYCERATE (2PG) (3PG)

DEHYDRATION OF 2PHOSPHOGLYCERATE
O C OThese 2 groups will detach to form water

H
H

C C H

P OH

2-PHOSPHOGLYCERATE (2PG) PHOSPHOENOL PYRUVATE (PEP)

Reversible Main product = Phosphoenol pyruvate (PEP) PEP is a high energy compound PEP has double bond Enzyme = Enolase

The phosphate group is replaced by carbonyl group

FORMATION OF PYRUVATE
Irreversible, large free energy drop Main product = Pyruvate ATP produced Pyuvate is one of the sources of acetyl CoA Enzyme = Pyruvate kinase (PK) Inhibitors of PK = ATP and acetyl CoA Activators of PK = fructose-1,6biphosphate Pyruvate transforms to Acetyl CoA in aerobic environment Pyruvate transforms to lactate (lactic acid) in anaerobic environment

O C

O-

C H C H

O P H

PHOSPHOENOL PYRUVATE PYRUVATE (PEP)

10
A P P P

O C

O-

C=O group is reduced back to OH (hydroxyl )

REDUCTION OF PYRUVATE TO LACTATE

Occurs when O2 supply is inadequate Reversible Main product = Lactate (lactic acid) Other product = NAD Reduction is performed by NADH (produced from rxn 6) as reducing agent Enzyme = Lactate dehydrogenase Lactate causes cramps and muscle pain Lactate is an end waste and thus eliminated from the cell and into general circulation (blood)

H
H

C C H

OH O
H

PYRUVATE (PYRUVIC ACID) LACTATE (LACTIC ACID)

11

Main regulator

Activated by AMP and fructose-2,3-biphosphate

Hexokinase and Glucokinase

They are enzymes used in phosphorylation of glucose When glucose is phosphorylated, it is practically trapped inside the cell (can no longer leak out) thus becomes committed to glycolysis. Glucokinase is present only in the liver while hexokinase is present in all tissues (including the liver) Hexokinase is inhibited by glucose-6-phosphate but not glucokinase. Glucokinase synthesis is activated by insulin but not hexokinase Hexokinase acts very rapidly but is easily exhausted (that is, easily saturated). Glucokinase acts more slowly but is not easily exhausted (high saturation)

Phosphofructokinase-1 (PFK-1)
PFK-1 is the main control valve for glycolysis It is activated by adenosine monophosphate (AMP) and fructose2,6-biphosphate (F2,6BP) It is deactivated (inhibited) by ATP and citrate. If it is inhibited, glycolysis slows down If it is activated, glycolysis speeds up. The most interesting activator of PFK-1 is Fructose-2,6-biphosphate (F2,6BP) because it activates glycolysis while deactivates gluconeogenesis. More F2,6BP = more active PFK-1 = faster glycolysis = deactivated gluconeogenesis Production of F2,6BP is governed by another enzyme called PKF-2 When PKF-2 has no phosphate group, it is active and activates the production of F2,6BP, increasing glycolysis. When PFK-2 is phosphorylated, it si deactivated so there are fewer F2,6BP so that gluconeogenesis is activated.

PYRUVATE KINASE (PK)

Enzyme used to convert phosphoenol pyrvate (PEP) to pyruvate with the corresponding production of ATP Inhibitors of PK = ATP and acetyl CoA Activators of PK = fructose-1,6-biphosphate When PK is deactivated (inhibited), phosphoenol pyruvate is transformed to oxaloacetate (OAA) instead of pyruvate. This is a sort of U turn for glycolysis, and the direction flows back to gluconeogenesis (an anabolic pathway of glucose Anabolic is the synthesis (building up) of a more complex product from simpler precursors. Gluconeogenesis, therefore, is the synthesis of glucose from noncarbohydrates precursors which are mostly glucogenic amino acids. Gluconeogenesis is very important in keeping the availability of glucose in the blood because some critical cells like RBC and brain cells use glucose exclusively (or almost exclusively) as their source of energy

1ST PHASE OF GLYCOLYSIS Dihydroxy 5

Acetone phosphate

NADH produced Glyceraldehyde -3-phosphate

2ND PHASE OF GLYCOLYSIS

1,3-biphospho glycerate

ATP produced

4
Fructose-1,6biphosphate ATP consumed Rate limiting step

Fructose-6phosphate

2
Glucose-6 phosphate ATP consumed

This is the product 3-phospho st phase of the 1 When O2 is glycerate and the raw adequate, all 8 material for the pyruvate 2nd phase. In the enters the 2-phospho 1st phase, 1 mitochondri glycerate glucose produces a where it is 2 of this, both of oxidized to 9 w/c will be Acetyl CoA Phosphoenol converted to to enter the pyruvate pyruvate in the 2nd Kreb cycle phase 10 ATP produced Rate limiting step Pyruvate NADH consumed Lactate

Glucose

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Fate of some pyruvate when O2 is inadequate

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