Kreb’s Cycle (aka, tricarboxylic acid (TCA)cycle, citric acid cycle

)
“The wheel is turnin’ and the sugar’s a burnin’”

Overall goal
• Makes ATP • Makes NADH • Makes FADH2 • Requires some carbohydrate to run

Geography
• Glycolysis in the cytosol • Krebs in mitochondrial matrix • Mitochondrion
– Outer membrane very permeable
• Space between membranes called intermembrane space (clever huh!)

– Inner membrane (cristae)
• Permeable to pyruvate, • Impermeable to fatty acids, NAD, etc

– Matrix is inside inner membrane

Conversion of pyruvate to Acetyl CoA
O H3C O pyruvate O
pyruvate dehydrogenase complex

NAD+

NADH

HSCoA

CO2 H3C

O SCoA

acetyl CoA

• • • •

2 per glucose (all of Kreb’s) Oxidative decarboxylation Makes NADH -33.4kJ

Fates of Acetyl CoA
O TAG's H3C SCoA Kreb's CO2, ATP, NADH...energy acetyl CoA

no CHO present

ketone bodies

• • •

In the presence of CHO an using energy
– Metabolized to CO2, NADH, FADH2,GTP and, ultimately, ATP

If energy not being used (Lots of ATP present)
– Made into fat

If energy being used, but no CHO present
– Starvation – Forms ketone bodies (see fat metabolism slides) – Danger!

Kreb’s Cycle
acetyl CoA SCoA H3C C O
O O
NADH C NAD HC OH CH2 malate
fumarase

O H2O

O

CoASH
HO

H2O

O

O

C
O

dehydrogenase

C C
H O

O

O

O C C O CH2 C O

+

citrate synthase

O

C CH2 O C C O CH2 citrate C
O

O

H

malate

oxaloacetate

aconitase

C C

fumarate
O

Kreb's Cycle
HO

O

succinate

FADH2 FAD O C CH2 CH2 GTP
SCoA O
NADH

dehydrogenase

O

CoASH GDP

C O O succinate

succinyl CoA synthetase

O

C CH2 CH2 CoASH C CO2
O
dehydrogenase

NAD

alpha ketoglutarate O O

C NADH C O CO2 CH2 CH2 C
O

C CH O HC C O CH2 isocitrate C
O O

O

NAD

isocitrate dehydrogenase

alpha ketoglutarate

succinyl CoA

O

Net From Kreb’s
• Oxidative process
– 3 NADH – FADH2 – GTP

• X 2 per glucose
– 6 NADH – 2 FADH2 – 2 GTP

• All ultimately turned into ATP (oxidative phosphorylation…later)

Citrate Synthase Reaction (First)
O
H3C C O SCoA

+

O

O C C O CH2 C O

O

H2O

CoASH
HO

citrate synthase
O

C CH2 O C C O CH2 C
O

O

acetyl CoA

oxaloacetate

citrate

• Claisen condensation • -32.2kJ

Aconitase Reaction
O HO

O

C CH2 O C C O CH2 C
O

O

O HO

aconitase

C CH HC C CH2 C
O O

O

O O

citrate

isocitrate

• Forms isocitrate • Goes through alkene intermediate (cis-aconitate)
– elimination then addition

• 13.3kJ

Isocitrate Dehydrogenase
O HO

C CH HC C CH2 C
O O

O

O
O O

NAD

NADH

CO2

C C O CH2 CH2 C
O

O

isocitrate dehydrogenase

O

isocitrate

alpha ketoglutarate

• All dehydrogenase reactions make NADH or FADH2 • Oxidative decarboxylation • -20.9kJ • Energy from increased entropy in gas formation

α-ketoglutarate dehydrogenase
O

C C O CH2 CH2 C
O

O

SCoA

O

CoASH

CO2

NAD

NADH

O

alpha ketoglutarate
dehydrogenase

O

C CH2 CH2 C
O

alpha ketoglutarate

succinyl CoA

• •

Same as pyruvate dehydrogenase reaction Formation of thioester
– endergonic – driven by loss of CO2
• increases entropy • exergonic

-33.5kJ

Succinyl CoA synthetase
SCoA O

O GDP GTP CoASH

O

C CH2 CH2 C
O

O C CH2

succinyl CoA

succinyl CoA synthetase

CH2 C O O succinate

• • •

Hydrolysis of thioester
– Releases CoASH – Exergonic

Coupled to synthesis of GTP
– Endergonic – GTP very similar to ATP and interconverted later

-2.9kJ

Succinate dehydrogenase
O C CH2 C O FAD FADH2
H
succinyl CoA
dehydrogenase

O

O

C C C C

H

CH2 O O

O

O

succinate

fumarate

• •

Dehydrogenation Uses FAD
– NAD used to oxidize oxygen-containing groups
• Aldehydes • alcohols

– FAD used to oxidize C-C bonds – 0kJ

Fumarase
O O O

O C HC OH CH2 C
O O

C C
H O

H2O
H
fumarase

C C

O

fumarate

malate

• Addition of water to a double bond • -3.8kJ

Malate Dehydrogenase
O

O C HC OH CH2 C
O O
NAD
NADH

O

dehydrogenase

malate

O

O C C O CH2 C O

malate

oxaloacetate

• • • •

Oxidation of secondary alcohol to ketone Makes NADH Regenerates oxaloacetate for another round 29.7 kJ

Net From Kreb’s
• Oxidative process
– 3 NADH – FADH2 – GTP

• X 2 per glucose
– 6 NADH – 2 FADH2 – 2 GTP

• All ultimately turned into ATP (oxidative phosphorylation…later)

Total Energy per glucose
• Cytosol
– Glycolysis
• 2 NADH • 2 ATP

• Mitochondrion
– Pyruvate dehydrogenase
• 2 NADH

• Krebs
– 6 NADH – 2 FADH2 – 2 GTP

Total Energy/glucose
• In mitochondrion:
– Each NADH makes 2.5 ATP – Each FADH2 makes 1.5 ATP – GTP makes ATP

• So…
– From in mitochondrion
• 8 NADH X 2.5 ATP/NADH = 20 ATP • 2 FADH2 X 1.5 ATP/FADH2= 3 ATP • 2 GTP X 1 ATP / GTP = 2 ATP • TOTAL in mitochondrion 25 ATP

Total Energy/ glucose
• Cytosol
– 2 ATP – 2 NADH
• NADH can’t get into mitochondrion • In eukaryotes two pathways,
– transferred to FADH2 » get 1.5 ATP/ FADH2 – Or transferred to NADH » Get 2.5 ATP/ NADH – (Not a problem in prokaryotes (why?))

– 2 NADH X 1.5 ATP – Or 2 NADH X 2.5 ATP
» + » Total

= 3 ATP = 5 ATP

=2 ATP 3+ 2 or 5 + 2 so either 5 or 7

ATP/glucose
• Eukaryotes
– – – – Mitochondrial: 25 ATP Cytosolic: 5 or 7 ATP Total 30 or 32 ATP/glucose 30 ATP X 7.3kcal X 4.18 kJ = 915 kJ
ATP If 32 ATP kcal = 976 kJ

• Prokaryotes
– 32 ATP X 7.3kcal X 4.18 kJ = 976 kJ
ATP kcal