THE TRICARBOXYLIC ACID CYCLE The TCA Cycle

 This process is aerobic, requiring

Introduction oxygen as the final oxidant of the
 TCA cycle is a series of reactions in mitochondria reducing equivalents
that bring about the  The enzymes of the citric acid cycle are
 Catabolism of acetyl residues located in the mitochondrial matrix
 Liberation of hydrogen equivalents  The 3 moles of NADH and 1 mole of
 Release and capture as ATP of most of FADH2 per round of the cycle  ETC
the available energy of tissue fuels  Each mole of NADH = 2.5
 The acetyl residues are in the form of acetyl-CoA moles of ATP
 CoA contains the vitamin pantothenic  FADH2 = 1.5 moles of ATP
acid  Each mole of pyruvate 
 Heterotrophic organisms TCA cycle, 10 moles of ATP
can be generated
 Obtain free energy by coupling their
metabolism to the breakdown of
complex organic molecules in their
environment
 ATP plays a central role in the
transference of free energy from
exergonic to endergonic processes
 Autotrophic organisms
 Use simple exergonic processes
 Energy of sunlight in plants
 The reaction Fe+2  Fe+3 in bacteria

Structure of a Mitochondrion Citrate Synthase

 Condensation of the methyl carbon of acetyl-CoA
with the keto carbon (C-2) of oxaloacetate (OAA)
 Excess citrate is used to transport acetyl-CoA
carbons from the mitochondrion to the cytoplasm
where they can be used for fatty acid and
cholesterol biosynthesis

Aconitase
 Aconitase is one of several mitochondrial enzymes
known as non-heme-iron proteins
 The isomerization of citrate to isocitrate by
aconitase is stereospecific

Isocitrate Dehydrogenase
 Isocitrate is oxidatively decarboxylated to -ketoglutarate by isocitrate dehydrogenase (IDH)
 IDH catalyzes the rate-limiting step, as well as the first NADH-yielding reaction of the TCA cycle

TCA is an integral part of the process by which much free
energy liberated during the oxidation of carbohydrates, lipids,
and amino acids is made available

During the course of oxidation of acetyl-CoA in the cycle,
reducing equivalents in the form of hydrogen or of electrons
are formed as a result of the activity of specific dehydrogenases
 subsequently being transferred to GDP
These reducing equivalents then enter the respiratory chain,
where large amounts of ATP are generated in the process of
oxidative phosphorylation
-Ketoglutarate Dehydrogenase Complex
 -ketoglutarate is oxidatively decarboxylated to succinyl-CoA by the -ketoglutarate
dehydrogenase (-KGDH) complex
 This reaction generates the second TCA cycle equivalent of CO2 and NADH
Succinyl-CoA Synthetase/Succinate Thiokinase
 The conversion of succinyl-CoA to succinate by succinyl-CoA synthetase involves the use of the
high-energy thioester of succinyl-CoA to drive the synthesis of a high-energy nucleotide
phosphate, by a process known as substrate-level phosphorylation
 In this process, a high energy enzyme-phosphate intermediate is formed with the phosphate
 Mitochondrial GTP is used in a trans-phosphorylation reaction catalyzed by the mitochondrial
enzyme nucleoside diphosphokinase to phosphorylate ADP, producing ATP and regenerating
GDP for the continued operation of succinyl-CoA synthetase
Succinate Dehydrogenase (SDH)  Substrate availability can also regulate TCA flux
 Succinate dehydrogenase catalyzes the oxidation of succinate to fumarate with the sequential  This occurs at the citrate synthase reaction as a result of reduced availability of
reduction of enzyme-bound FAD and non-heme iron oxaloacetate
 In mammalian cells, the final electron acceptor is coenzyme Q (CoQ), a mobile carrier of  Product inhibition also controls the TCA flux
reducing equivalents that is restricted by its lipophilic nature to the lipid phase of the  Citrate inhibits citrate synthase
mitochondrial membrane  -ketoglutarate dehydrogenase is inhibited by NADH and succinyl-CoA
 The key enzymes of the TCA cycle are also regulated allosterically by Ca2+, ATP, and ADP
Fumarase (Fumarate Hydratase)
 The fumarase-catalyzed reactions are specific for the trans form of fumarate
 The result is that the hydration of fumarate proceeds stereospecifically with the production of
L-malate

Malate Dehydrogenase (MDH)

 L-malate is the specific substrate for MDH, the final enzyme of the TCA cycle
 The forward reaction of the cycle, the oxidation of malate, yields oxaloacetate (OAA)

10 ATP are Formed per Turn of the TCA Cycle

Reaction Catalyzed by Method of Production ATP Molecules Formed

Isocitrate dehydrogenase Respiratory chain oxidation of NADH 2.5
-Ketoglutarate dehydrogenase Respiratory chain oxidation of NADH 2.5
Succinate thiokinase Phosphorylation at substrate level 1
Succinate dehydrogenase Respiratory chain oxidation of FADH2 1.5
Malate dehydrogenase Respiratory chain oxidation of NADH 2.5

Regulation of the TCA Cycle

 Regulation of the TCA cycle, like that of glycolysis, occurs at both the level of entry of substrates
into the cycle as well as at the key reactions of the cycle
 Fuel enters the TCA cycle primarily as acetyl-CoA
 The generation of acetyl-CoA from carbohydrates is, therefore, a major control point
of the cycle
 This is the reaction catalyzed by the pyruvate dehydrogenase (PDH) complex
Pyruvate Dehydrogenase Complex
Role in Metabolism
 The TCA cycle plays roles in both oxidative and synthetic processes; it is amphibolic
 The TCA cycle takes part in gluconeogenesis, transamination and deamination, and fatty acid
synthesis
 All major members of the cycle are potentially glucogenic since they give rise to a net
production of glucose in the liver or kidney, the organs that contain a complete set of enzymes
necessary for gluconeogenesis
 Aminotransferase reactions produce pyruvate from alanine, oxaloacetate from aspartate, and
-ketoglutarate from glutamate
 Because these reactions are reversible, the cycle also serves as a source of carbon skeletons for
the synthesis of nonessential amino acids

TCA Intermediates and Metabolism

 Since three reactions of the TCA cycle as well as PDH utilize NAD + as a co-factor, it is not
difficult to understand why the cellular ratio of NAD+/NADH has a major impact on the flux of
carbon through the TCA cycle