Function

Citrate Synthesis
The enzyme citrate synthase catalyzes the formation of citrate from acetyl CoA and
oxaloacetate, often regarded as the first step of the TCA cycle. This reaction is virtually
irreversible and has a delta-G-prime of -7.7 Kcal/M that strongly favors citrate formation.
The availability of substrates and products regulates the activity of citrate synthase. For
instance, citrate itself acts as an inhibitor for citrate synthase, while oxaloacetate’s binding
to it increases its affinity for acetyl-CoA. It bears mention that phosphofructokinase-1 in
glycolysis is inhibited by citrate, while it activates acetyl-CoA carboxylase for fatty acid
synthesis. This point illustrates the interconnectivity of our metabolic cycles.[5]
Isomerization of Citrate
The reversible conversion of citrate to isocitrate is catalyzed by the enzyme aconitase,
which contains an iron-sulfur center that facilitates the hydroxyl group migration. Cis-
aconitate is the intermediate product of this reaction.[6]
Oxidative Decarboxylation of Isocitrate
The oxidative decarboxylation of isocitrate to alpha-ketoglutarate becomes catalyzed by
NAD+ -dependent isocitrate dehydrogenase producing CO2, NADH, and a proton; this is
the rate-limiting step of the TCA cycle. Production of the first reduced coenzyme in the
cycle takes place at this reaction. The tendency of this reaction to produce gas makes it
irreversible. ADP and calcium ions allosterically regulate isocitrate dehydrogenase by
activating it, while ATP and NADH inhibit its activity.[7]
Oxidative Decarboxylation of Alpha-ketoglutarate
The conversion of alpha-ketoglutarate to succinyl-CoA is catalyzed by the alpha-
ketoglutarate dehydrogenase complex producing NADH, CO2, and H+. The function of the
alpha-ketoglutarate dehydrogenase complex is analogous to PDC. Alpha-ketoglutarate
becomes decarboxylated by E1 of this complex, transferring the four remaining carbons to
thiamine pyrophosphate. Thiamine pyrophosphate is the first cofactor. Then the succinyl
group transfers to CoASH by E2 with the help of FAD. The final step involves the
resynthesis of FAD along with NADH from NAD+ by E3. This last step ensures the
maintenance of substrates and cofactors required to continue the dehydrogenase complex
activity. The cofactors necessary for alpha-ketoglutarate dehydrogenase complex include
thiamine pyrophosphate, lipoic acid, coenzyme A, NAD+, and FAD. Succinyl-CoA,
NADH, and ATP inhibit the alpha-ketoglutarate dehydrogenase complex.[8][9]
Cleavage of Succinyl Coenzyme A
The enzyme succinate thiokinase catalyzes the reversible interconversion of succinyl-CoA
to succinate by cleaving succinyl CoA’s thioester bond. The enzyme uses inorganic
phosphate and dinucleotide to produce trinucleotide and CoA. This coupled reaction is
called substrate-level phosphorylation, just like what happens in glycolysis.[10]
Oxidation of Succinate
Succinate dehydrogenase is also called complex II due to its role in the aerobic respiration
chain. It catalyzes the reduction of ubiquinone to ubiquinol. The TCA cycle catalyzes the
oxidation of succinate to fumarate, producing a reduced FADH2 from FAD.[11]
Hydration of Fumarate
The reversible hydration of fumarate to malate is catalyzed by fumarase (or fumarate
hydratase). In another attempt to illustrate the interconnectedness of metabolic pathways,
note that fumarate production also occurs in the urea cycle.[12]
Oxidation of Malate
Malate dehydrogenase is the catalyst in the reversible oxidation of malate to oxaloacetate,
which is the last step of the TCA cycle. This enzyme plays a crucial role in NADH
oxidation within the TCA cycle. The delta-G-prime is positive, which indicates that the
reaction favors malate. However, the consumption of oxaloacetate by citrate synthase
drives this reaction forward to produce more oxaloacetate.[13]
Cataplerotic Processes
Citric acid intermediates can leave the cycle to participate in the biosynthesis of other
compounds. Citrate can be directed toward fatty acids; succinyl-CoA to heme synthesis;
alpha-ketoglutarate to amino acid synthesis, purine synthesis, and neurotransmitter
synthesis; oxaloacetate to amino acid synthesis, and malate to gluconeogenesis.[14][4]
Anaplerotic Processes
Intermediates are inserted into the TCA cycle to replace the cataplerotic processes and
ensure the continuation of the cycle. For instance, pyruvate can enter the cycle throughout
the body through pyruvate carboxylase, which inserts additional oxaloacetate into the cycle.
This process causes the reaction to be pushed forward toward the already exergonic citrate
synthase, as there is more oxaloacetate to participate in the reaction. The liver is another
example, as it can produce alpha-ketoglutarate by oxidative deamination or transamination
of glutamate.[15][4]