The carbon sequence is oxaloacetate plus an acetyl group to citrate , then isocitrate , alpha-ketoglutarate , succinyl-CoA , succinate , fumarate , malate and back to oxaloacetate . The numbers count the cycling carbon skeleton, excluding coenzyme A. Two carbon dioxide releases accompany the changes .[ref]
Figure 1.
Carbon counts and electron carrier outputs around the citric acid cycle
. Original schematic of one forward turn. The carbon skeleton excludes coenzyme A. Labels show NADH, carbon dioxide, GTP and ubiquinol outputs. Positions and arrow lengths carry no structural or kinetic meaning. Details.
Per acetyl group: three NADH, one FADH2 equivalent, one GTP or ATP by substrate-level phosphorylation, and two carbon dioxide. Succinate dehydrogenase retains FADH2 and transfers its Electrons to ubiquinone, forming ubiquinol. The FADH2 equivalent and ubiquinol count the same Electron pair.[ref]
A useful check adds the stepwise changes. The carbon count is ; the NADH count is . Each NADH carries two Electrons, and the succinate dehydrogenase route supplies another pair, so one turn exports Electrons. This arithmetic is an accounting identity, not an ATP yield calculation. Converting it to ATP requires the separate energy coupling and transport assumptions discussed under oxidative phosphorylation.
For a chosen ideal conversion of of acetyl groups, multiplication gives of NADH, of ubiquinol, of GTP and of carbon dioxide. The Electron output is . If all these Electrons subsequently reach oxygen with no competing sinks, four Electrons per oxygen molecule require of oxygen. That is consumption by the connected electron transport chain; it is not an oxygen term in the cycle's own substrate ledger.
No cycle step directly consumes molecular oxygen. In ordinary aerobic mitochondrial operation, however, the mitochondrial respiratory chain must reoxidize NADH and ubiquinol to restore the oxidized electron carriers. Without that outlet, a finite oxidized-carrier pool cannot sustain continued turnover. Other electron acceptors and other forms of metabolism require their own analysis.[ref]
The pyruvate dehydrogenase complex acts upstream: one three-carbon pyruvate gives one two-carbon acetyl group, one carbon dioxide and one NADH. These are additional to the cycle ledger.[ref] Consequently, processing one pyruvate through both stages gives three carbon dioxide and four NADH, while the succinate dehydrogenase contribution and the substrate-level phosphorylation contribution each remain one. Counting the upstream step inside the cycle would obscure where the extra carbon dioxide and NADH arise.
Net stoichiometry does not identify individual chemical atoms. In the ordinary forward citric acid cycle, neither entering acetyl group carbon is released in that same first turn: both released carbon dioxide carbon chemical atoms came from the starting oxaloacetate. The entering carbon remains in regenerated intermediates.[ref]
Consider a chosen ideal first-turn experiment, with negligible natural carbon-13 abundance and no exchange, side pathways, repeated turns or isotope effects. Supply of initially unlabelled oxaloacetate and of acetyl-CoA, of which has both acetyl group carbon positions labelled with carbon-13. The other has neither position labelled. The label refers only to the acetyl group, not to coenzyme A.
The input contains of labelled carbon chemical atoms. The of citrate initially formed contains of carbon chemical atoms, so its labelled-carbon fraction is . After exactly one forward traversal per starting oxaloacetate, the of released carbon dioxide has zero label, while the regenerated of oxaloacetate contains all of labelled carbon chemical atoms among total: .
Figure 1.
First-turn carbon-13 accounting distinguishes net stoichiometry from chemical atom identity
. Original ideal calculation, not historical or measured isotope data. Stacked bars show the amount of substance of carbon chemical atoms, not amounts of whole molecules. The first two bars are alternative stages; the last two are simultaneous outputs whose totals sum to the preceding citrate bar. Carbon-13 is restricted initially to the incoming acetyl groups; natural abundance and side pathways are excluded. Details.
The conservation check is for total carbon and for label. In this model, of regenerated oxaloacetate molecules carry label, distinct from the labelled-carbon fraction. The regenerated oxaloacetate is the same chemical species and the same amount of substance as the initial acceptor pool, but its chemical atom identities have changed. Saying that the cycle oxidizes an acetyl group to two carbon dioxide is a net ledger, not a claim that those exact two incoming chemical atoms immediately leave. Later-turn label release requires positional tracking and mixing assumptions; the first-turn balance alone does not determine it.