Glycolysis

Chapter 16.1-16.2 The first stages of metabolizing sugars anaerobically

TCA Cycle

Anaerobic and Aerobic Metabolism of Glucose

The Glycolytic pathway splits a six carbon sugar into two 3-carbon compounds in three stages

It is common to almost all organisms

The First Stage of Glycolysis

Stage 1

Hexokinase uses Induced Fit

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Hexokinase Uses an Induced Fit

E + Glucose ES

Voet and Voet, Wiley

Preparing for the split

Isomerase

Aldolase is the enzyme that cleaves the critical bond

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Aldolase reaction - retroaldol condensation

R R

O + R-NH3

R R N R H

H2O

Schiff Base

See pg 606 (582) for details

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Stage 2: Isomerization of Dihydroxyacetone phosphate to glyceraldehyde 3-phosphate

Triose phosphate isomerase interconverts dihydroxyacetone phosphate and glyceraldehyde 3phosphate Two molecules of glyceraldehyde 3-phosphate can now enter Stage 3 of glycolysis only one path to finish 12

II

III

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Triose phosphate isomerase (TIM)

kcat/Km = 2 x 108 M-1s-1 !!!

TIM-barrel ,-Barrel Very stable and common

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TIM Mechanism

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Stage III is Productive!

No gain of energy 2 ATP

Here is the reward! (2 ATP) x 2 + 2 NADH

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II

III

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Glyceraldehyde-3-P Dehydrogenase

This uses the favorable oxidation of an aldehyde to an acid to obtain NADH and a high energy phosphate - an anhydride 18

Glyceraldehyde 3- phosphate dehydrogenase oxidizes and phosphorylates with one enzyme

Fig. 16.8, Stryer
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Energetics of 1,3 Bisphosphoglycerate Formation

- 12 kcal/mol

+ 12 kcal/mol

Formed with an active site cysteine residue during catalysissee Stryer, Figure 16.8
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II

III

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Phosphoglycerate Kinase generates the first ATP

Acid phosphatean anhydride

1,3 BPG has a high phosphotransfer potential: G = -11.8 kcal/mole (hydrolysis) In comparison, for ATP s phosphotransfer potential: G = -7.3 kcal/mole (hydrolysis)

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Generating the last ATP molecule: three enzymatic steps

Phosphoenolpyruvate has a high phosphotransfer potential:

G = -14.8 kcal/mole (hydrolysis)
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Note that the G values in the last column, which are calculated for typical concentrations of these compounds in the cell, are much more favorable than the G

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Fate of Pyruvate

TCA Cycle More energy

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Anaerobic Fermentation + Regenerates NAD

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Entry of galactose and fructose into the glycolytic pathway

Read pgs 469-472 to see how these interconversions take place

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The energy charge of a cell regulates catabolic and anabolic reactions

Normal energy charge

Energy Charge = [ATP] +1/2[ADP] [ATP]+[ADP]+[AMP] Adenylate kinase AMP+ATP <=> ADP+ADP

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Regulation of phosphofructokinase is key to regulating glycolysis

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Regulation of phosphofructokinase is key to regulating glycolysis

30 Inhibition of phosphofructokinase by binding of ATP at allosteric regulatory sites

Regulation of Glycolysis

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Glycolysis Regulation

32 See Pg 454, Stryer

Regulation of Pyruvate Kinase:

Allosteric and covalent modification (16.2)

Phosphorylation

Allosteric
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Glycolysis

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Glycolysis - Summary
Common to most organisms NADH must be converted to NAD+ by various means Glycolysis is tightly regulated in multiple ways Regulation is at irreversible steps

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