Citric Acid Cycle

What is it?
Series of rxns that oxidize acetyl CoA to 2CO2 in a manner that
conserves the liberated free energy for ATP production
Breakdown products of glc, fats and amino acids are all oxidized via
the cycle
Intermediates also a source for many biosynthetic pathways amphibolic which means serves in both catabolic and anabolic
processes

Also called TCA cycle - tricarboxylic acid cycle or Krebs cycle

Citric Acid Cycle

Cellular respiration

Citric Acid Cycle

Pyruvate dehydrogenase complex
3 mitochondrial euk enzymes
5 coenzymes

Citric Acid Cycle

Pyruvate dehydrogenase complex
5 coenzymes - Coenzyme A (pantothenate)
Reactive thiol is an acyl carrier

Citric Acid Cycle

Pyruvate dehydrogenase complex
5 coenzymes - NAD+ (niacin)
Involved in redox, electron carrier

Citric Acid Cycle

Pyruvate dehydrogenase complex
5 coenzymes - FAD (riboflavin, vitamin B2)
Involved in redox, electron carrier

Citric Acid Cycle

Pyruvate dehydrogenase complex
5 coenzymes - TPP (thiamine, vitamin B1)
Helps in cleavage of bonds next to carbonyl

Citric Acid Cycle

Pyruvate dehydrogenase complex
5 coenzymes - lipoate
Reversible oxidation at thiols, serves as electron carrier and acyl
carrier

Citric Acid Cycle

Pyruvate dehydrogenase complex

Citric Acid Cycle

Pyruvate dehydrogenase complex

Citric Acid Cycle

Pyruvate dehydrogenase complex - regulation
Eukaryotic complex regulated by phosphorylation

Regulation
Pyruvate: substrate for complex
ADP: indicates low energy charge
NADH, acetyl CoA: endproducts of complex
Ca2+: certain hormones (insulin) raise [Ca2+]

Citric Acid Cycle

Every turn of cycle:
1. Joining an acetyl group to oxaloacetate
2. Oxidizing 2 carbons off as CO2, leaving succinate
3. Converting succinate back to oxaloacetate
4. Producing 1 GTP and reduced cofactors

Citric Acid Cycle

Net reaction:

AcetylCoA + 3NAD+ + FAD + GDP + Pi + 2H2O 2CO2 + CoA + 3NADH + FADH2 +

GTP + 2H+

Citric Acid Cycle

Net reaction:

AcetylCoA + 3NAD+ + FAD + GDP + Pi + 2H2O 2CO2 + CoA + 3NADH + FADH2 +

GTP + 2H+

Citric Acid Cycle

Goal:
capture energy from acetate 2CO2
1. One GTP made: equivalent to 1 ATP
2. Lots of reduced cofactors, reducing power can be converted to ATP

-G irreversible

-G irreversible
+G
made favorable by fast
product removal

-ketoglutarate
dehydrogenase
complex
-G irreversible

Only enzyme bound to

inner mitochondrial membrane

Citric Acid Cycle

Regulation
Rate of cycle is determined by:
1. Availability of substrates (acetyl CoA, NAD+, FAD)
2. Inhibition by accumulating products
3. Allosteric feedback inhibition of enzymes

NAD+ and FAD are regenerated

by oxidative phosphorylation,
which needs O2

Citric Acid Cycle

Regulation
Three nonequilibrium rxns:
1. Citrate synthase
2. Isocitrate dehydrogenase
3. -ketoglutarate dehydrogenase

Citric Acid Cycle

Why so complicated?
Hub of intermediary metabolism
Greatest selective advantage

Citric Acid Cycle

Citric Acid Cycle

Reaction 1 - Citrate synthase
Condensation of acetylCoA with oxaloacetate to form citrate
Irreversible

Citric Acid Cycle

Reaction 1 - Citrate synthase
Condensation of acetylCoA with oxaloacetate to form citrate
Irreversible

Citric Acid Cycle

Reaction 2 - Aconitase
Removal of H2O, stereospecifically add back H2O
Reversible, pulled to right because isocitrate rapidly consumed

Prochiral carbon allows for stereospecific rxn

Citric Acid Cycle

Reaction 2 - Aconitase
Removal of H2O, stereospecifically add back H2O
Reversible, pulled to right because isocitrate rapidly consumed

Citric Acid Cycle

Reaction 3 - isocitrate dehydrogenase
Oxidative decarboxylation of isocitrate
Irreversible

Citric Acid Cycle

Reaction 4 - -ketoglutarate dehydrogenase complex
Oxidative decarboxylation of -ketoglutarate
Identical to pyruvate dehydrogenase reaction
Three homologous enzymes (E1, E2, E3)
Five coenzymes (TPP, lipoate, FAD, NAD+, CoA)
Irreversible

Citric Acid Cycle

Reaction 5 - Succinyl-CoA synthetase
Hydrolysis of thioester bond energizes formation of GTP
Synthetase indicates participation of nucleoside triphosphate
Reversible

Citric Acid Cycle

Reaction 6 - Succinate dehydrogenase
Oxidation of succinate to fumarate
Enzyme bound to inner mito membrane, only membrane-bound enz
Reversible

Malonate (analog of succinate) is a competitive inhibitor of enz,

blocks activity of cycle

Citric Acid Cycle

Reaction 7 - Fumarase
Hydration of fumarate to L-malate
Reversible

Substrate

Product

Citric Acid Cycle

Reaction 8 - Malate dehydrogenase
Oxidation of malate to oxaloacetate
NAD+ linked to enzyme
High +G, pulled to products by continual removal of oxaloacetate

Citric Acid Cycle

Defects in TCA

Mutant isocitrate dehydrogenase

gains new activity production of
2-hydroxyglutarate
High levels of 2-hydroxyglutarate
inhibit histone demethylase,
altering gene expression (seen in
glial cell tumors in brain)