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Lecture 14 10/3/05

Cellular Respiration:

Harvesting Chemical Energy

Chapter 9





II.

Catabolic

Oxidative

Pathways





1

Lecture Outline



1. Review Fatty Acid Oxidation Pathway

2. Oxidation of Monosaccharides (Glycolysis Pathway)

3. Mitochondrial Spaces: Matrix, Intermembrane Space

4. Oxidation of Pyruvate

5. Complete oxidation of Carbon from FA ox & glycolysis

– TCA Cycle (Kreb’s Cycle)

6. Cashing in the “accumulated” Reducing Equivalents -

Oxidative Phosphorylation









2

Fatty Acid Oxidation (b-oxidation) in Mitochondria

Priming Saturated

Step hydrocarbon





2e-

2 H+

-steps down removed

Ester

oxidation (acid)

states of carbon unsaturated

hydrocarbon



-captures

Reducing potential

NADH + H+

FADH2 Ketone

2e-

alcohol

2 H+ 3

removed

Net Result of Fatty Acid Oxidation Pathway



Fatty acid shortened by 2 carbon unit



2 carbon acid attached to CoA (acetyl CoA)



Oxidation of Carbon -CH2- to –C=O

to acid

S CoA

Capture reducing equivalents

2 NADH + H+

2 FADH2

4

Glycolysis Another catabolic pathway

Oxidation of monosaccharides

(polysaccharides digested to monosaccharides; no energy harvested)



Hexoses (like glucose)

6 carbon sugar

Aldehyde with 5 alcohols









(Two)

3 carbon acid

with a ketone

oxidized to pyruvic acid

5

Glycolysis consists of two major phases

Energy investment phase







Glucose



Energy investment phase

(Priming Steps) 2 ATP + 2 P 2 ATP used









Energy payoff phase









4 ADP + 4 P 4 ATP formed









Energy payoff phase

2 NAD+ + 4 e- + 4 H + 2 NADH + 2 H+









2 Pyruvate + 2 H2O









Net Harvest Glucose 2 Pyruvate + 2 H2O







Of Pathway 4 ATP formed – 2 ATP used 2 ATP + 2 H+





2 NADH

6

2 NAD+ + 4 e– + 4 H +

Figure 9.8

CH2OH





Glycolysis

H H

H

HO H

HO OH

H OH





Energy Priming Stage

Glucose



ATP 1



Hexokinase

ADP





Add Pi to #6 carbon CH2OH

H

H

P

O H

OH H

HO

H OH

Glucose-6-phosphate





Rearrange

2

Phosphoglucoisomerase

CH2O P

O CH2OH

H HO

H HO

HO H

Fructose-6-phosphate







Add Pi to #1 carbon

3

ATP

Phosphofructokinase



ADP



P O CH2 O CH2 O P





Split into two

HO

H OH

HO H



2 of

3 carbon units

Fructose-

1, 6-bisphosphate

4





these

Aldolase



5 H

P O CH2 Isomerase C O





rearrange

C O

CHOH

CH2OH

CH2 O P

Dihydroxyacetone 7

Glyceraldehyde-

phosphate 3-phosphate

Figure 9.9 A

Glycolysis Two

6

2 NAD+

Triose phosphate

dehydrogenase









Energy PAYOFF Stage Of

2 NADH 2 Pi

+ 2 H+

2

P O C O







these

CHOH



CH2 O P





Oxidize #1 aldehyde to Acid

1, 3-Bisphosphoglycerate

2 ADP

7





NADH + H+, high energy phosphate

Phosphoglycerokinase



2 ATP





2 O–





Harvest Phosphate C

CHOH





Substrate Level Phosphorylation CH2 O

3-Phosphoglycerate

P









ATP made

8

Phosphoglyceromutase







rearrange

2 O–



C O

H C O P



CH2OH

2-Phosphoglycerate

9





Oxidize #2 alcohol to Ketone 2 H2O Enolase



2 O–





High energy phosphate

C O

C O P



CH2

Phosphoenolpyruvate

2 ADP

10

Pyruvate kinase







Harvest Phosphate pyruvate

2 ATP





2 O–





Substrate Level Phosphorylation C



C

O

O

8

ATP made Figure 9.8 B

CH3

Pyruvate

Net Result of Glycolysis Pathway



6 Carbon Sugar oxidized to

TWO Pyruvate (acid with ketone)

Net yield of 2 ATP produced

by substrate level phosphorylation



Capture reducing equivalents

2 NADH + H+



9

Fate of the 2 Pyruvate molecules?



enter the mitochondrion for further oxidation

produce 6 CO2



Pathway called the TCA cycle







What does it mean to “get into” the

mitochondrion?



Glycolysis occurs in cytosol 10

Mitochondria

Functional Spaces



Matrix

Inner

Mito

Membrane





Outer

Mitochondrial

Folds of

Inner membrane Membrane

Called

Cristae Intermembrane

Space 11

Electrons Electrons carried

carried via NADH and

via NADH FADH2









Oxidative

Glycolsis Citric phosphorylation:

acid electron transport

Glucose Pyruvate cycle and

chemiosmosis





Cytosol

Mitochondrion









ATP ATP ATP



Substrate-level

Substrate-level Oxidative

phosphorylation

phosphorylation phosphorylation

Figure 9.6

12

Matrix

Contains DNA

Contains bacterial-like ribosomes

Site of three important oxidative reaction cycles

Fatty Acid Oxidation

Pyruvate oxidation

TCA Cycle



Intermembrane Space

Reservoir to hold H+ ions



Inner Mitochondrial Membrane

Densely packed with proteins

Site of “oxidative phosphorylation”

electron transport and ATP production13

Oxidation of Pyruvate to Acetyl CoA

Familiar?

CYTOSOL MITOCHONDRION









NAD+ NADH + H+



O–

S CoA

2

C O

C O



C O



CH3

1 3







Acetyl

CH3

Acetyl CoA



pyruvate

Pyruvate CO2 Coenzyme A





CoA

Transport protein

14

Figure 9.10

Digestion

Proteins Carbohydrates Fats









Amino Sugars Glycerol Fatty

acids acids









Glycolysis



Glucose

Catabolic

Glyceraldehyde-3- P

Pathways

NH3 Pyruvate







Acetyl CoA









Interconnected

Pathways

Citric

acid

cycle









Common

Figure 9.19

Oxidative

phosphorylation Intermediates

15

Pyruvate

Oxidation





Tri Carboxylic Acid

TCA Cycle

Kreb’s Cycle









16

TCA Cycle

Stepping down the Oxidation Series of Carbons

-Acid to CO2

-Ketone to acid (thioester)

- saturated to unsaturated

- unsaturated to alcohol

- alcohol to ketone or carboxylic acid

- decarboxylation



Capture reducing equivalents

NAD+ 2e- + 2H+ NADH + H+

FAD 2e- + 2H+ FADH2

17

TCA Cycle Pyruvate

(from glycolysis,

Glycolysis Citric

acid

cycle

Oxidative

phosphorylation



2 molecules per glucose)



•Oxidize the 2 carbon unit ATP ATP ATP









Acetyl CoA CO2







To 2 CO2

CoA

NADH







•Five key oxidation steps

+ 3 H+ Acetyle CoA

CoA





3 NADH + H+ CoA







1 FADH2

1 GTP Citric



Per 2 carbon unit acid

cycle

2 CO2





FADH2 3 NAD+









Regenerate the initial

FAD 3 NADH



+ 3 H+





4 carbon acid

ADP + P i



ATP



repeat

18

Figure 9.11

Reactions of the TCA cycle pathway



Note oxidation points

Capture of 3 NADH + H+

Capture of 1 FADH2

Capture of 1 high energy phosphate bond (GTP)

per 2 carbon unit



So some GTP harvested,

but a lot of reducing equivalents accumulated

these are worth a lot more ATP



How? 19

What good are “accumulated” reducing equivalents?

H2 + 1/2 O2









H2 = 2e- 2H+









Free energy, G

Explosive

release of

heat and light

energy







2H2 + O2 = H2O + BOOM!

Figure 9.5 A H2O









20

The Regeneration Energy Carriers

Energy carriers (ATP, NAD+, FAD) present

in only minute amounts

Only limited amounts of reducing equiv carriers

must “cash them in” - oxidize them

2e-

2e- 2H+

2H+ Cashed in

Captured in catabolism





Electron transport

– oxidizes NADH and FADH2 NADH + H+

back to NAD+ and FAD









Energy from catabolism Energy for cellular work

(exergonic, energy yielding (endergonic, energy-

processes) consuming processes)

NAD+ 21

Mitochondrial Functions



Oxidize compounds to CO2 + H2O

Fatty acid Oxidation

Produce

Oxidation of Pyruvate reduced carriers

TCA Kreb’s Cycle NADH & FADH2

Generate >90% of Typical Cell’s ATP

Oxidative Phosphorylation

“electron transport”

ATP synthesis Oxidize reduced carriers

to produce ATP or equiv

22

Next time



How cash in the NADH + H+ and FADH2

“poker chips” for ATP



Oxidative Phosphorylation









23

Summary

-FA oxidation in matrix of mitochondrion

-Glycolysis (ox of sugar) in cytosol

-Oxidation of pyruvate in matrix of mitochondrion

-TCA cycle (oxidation of acetyl CoA) in matrix



- “accumulate” reducing equivalent carriers

- must “cash in” for ATP – oxidative Phosphorylation

Electrons Electrons carried

carried via NADH and

via NADH FADH2





Citric

Glycolsis acid Oxidative

Glucose Pyruvate phosphorylation

cycle



Cytosol Mitochondrion





ATP ATP ATP

Substrate-level 24

Substrate-level Oxidative

phosphorylation phosphorylation phosphorylation

Figure 9.6



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