Proline Oxidation Supports Mitochondrial ATP Production When Complex I Is Inhibited

Int J Mol Sci. 2022 May 4;23(9):5111. doi: 10.3390/ijms23095111.

Abstract

The oxidation of proline to pyrroline-5-carboxylate (P5C) leads to the transfer of electrons to ubiquinone in mitochondria that express proline dehydrogenase (ProDH). This electron transfer supports Complexes CIII and CIV, thus generating the protonmotive force. Further catabolism of P5C forms glutamate, which fuels the citric acid cycle that yields the reducing equivalents that sustain oxidative phosphorylation. However, P5C and glutamate catabolism depend on CI activity due to NAD+ requirements. NextGen-O2k (Oroboros Instruments) was used to measure proline oxidation in isolated mitochondria of various mouse tissues. Simultaneous measurements of oxygen consumption, membrane potential, NADH, and the ubiquinone redox state were correlated to ProDH activity and F1FO-ATPase directionality. Proline catabolism generated a sufficiently high membrane potential that was able to maintain the F1FO-ATPase operation in the forward mode. This was observed in CI-inhibited mouse liver and kidney mitochondria that exhibited high levels of proline oxidation and ProDH activity. This action was not observed under anoxia or when either CIII or CIV were inhibited. The duroquinone fueling of CIII and CIV partially reproduced the effects of proline. Excess glutamate, however, could not reproduce the proline effect, suggesting that processes upstream of the glutamate conversion from proline were involved. The ProDH inhibitors tetrahydro-2-furoic acid and, to a lesser extent, S-5-oxo-2-tetrahydrofurancarboxylic acid abolished all proline effects. The data show that ProDH-directed proline catabolism could generate sufficient CIII and CIV proton pumping, thus supporting ATP production by the F1FO-ATPase even under CI inhibition.

Keywords: coenzyme Q; proline dehydrogenase; reducing equivalent; substrate-level phosphorylation.

MeSH terms

  • Adenosine Triphosphatases / metabolism
  • Adenosine Triphosphate / metabolism
  • Animals
  • Electron Transport Complex I / metabolism
  • Glutamic Acid / metabolism
  • Mice
  • Mitochondria / metabolism
  • Proline / metabolism
  • Proline Oxidase* / metabolism
  • Ubiquinone* / metabolism

Substances

  • Ubiquinone
  • Glutamic Acid
  • Adenosine Triphosphate
  • Proline
  • Proline Oxidase
  • Adenosine Triphosphatases
  • Electron Transport Complex I