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. 1995 Nov;109(3):829–837. doi: 10.1104/pp.109.3.829

Electron Partitioning between the Cytochrome and Alternative Pathways in Plant Mitochondria.

M Ribas-Carbo 1, J A Berry 1, D Yakir 1, L Giles 1, S A Robinson 1, A M Lennon 1, J N Siedow 1
PMCID: PMC161383  PMID: 12228636

Abstract

The contribution of the cyanide-resistant, alternative pathway to plant mitochondrial electron transport has been studied using a modified aqueous phase on-line mass spectrometry-gas chromatography system. This technique permits direct measurement of the partitioning of electrons between the cytochrome and alternative pathways in the absence of added inhibitors. We demonstrate that in mitochondria isolated from soybean (Glycine max L. cv Ransom) cotyledons, the alternative pathway contributes significantly to oxygen uptake under state 4 conditions, when succinate is used as a substrate. However, when NADH is the substrate, addition of pyruvate, an allosteric activator of the alternative pathway, is required to achieve the same level of alternative pathway activity. Under state 3 conditions, when the reduction state of the ubiquinone pool is low, the addition of pyruvate allows the alternative pathway to compete with the cytochrome pathway for electrons from the ubiquinone pool when the cytochrome pathway is not saturated. These results provide direct experimental verification of the kinetics consequences of pyruvate addition on the partitioning of electron flow between the two respiratory pathways. This distribution of electrons between the two unsaturated pathways could not be measured using conventional oxygen electrode methods and illustrates a clear advantage of the mass spectrometry technique. These results have significant ramifications for studies of plant respiration using the oxygen electrode, particularly those studies involving intact tissues.

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Selected References

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  1. Bahr J. T., Bonner W. D., Jr Cyanide-insensitive respiration. II. Control of the alternate pathway. J Biol Chem. 1973 May 25;248(10):3446–3450. [PubMed] [Google Scholar]
  2. Day D. A., Dry I. B., Soole K. L., Wiskich J. T., Moore A. L. Regulation of Alternative Pathway Activity in Plant Mitochondria : Deviations from Q-Pool Behavior during Oxidation of NADH and Quinols. Plant Physiol. 1991 Mar;95(3):948–953. doi: 10.1104/pp.95.3.948. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Day D. A., Millar A. H., Wiskich J. T., Whelan J. Regulation of Alternative Oxidase Activity by Pyruvate in Soybean Mitochondria. Plant Physiol. 1994 Dec;106(4):1421–1427. doi: 10.1104/pp.106.4.1421. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Dry I. B., Moore A. L., Day D. A., Wiskich J. T. Regulation of alternative pathway activity in plant mitochondria: nonlinear relationship between electron flux and the redox poise of the quinone pool. Arch Biochem Biophys. 1989 Aug 15;273(1):148–157. doi: 10.1016/0003-9861(89)90173-2. [DOI] [PubMed] [Google Scholar]
  5. Elthon T. E., McIntosh L. Identification of the alternative terminal oxidase of higher plant mitochondria. Proc Natl Acad Sci U S A. 1987 Dec;84(23):8399–8403. doi: 10.1073/pnas.84.23.8399. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elthon T. E., Nickels R. L., McIntosh L. Monoclonal antibodies to the alternative oxidase of higher plant mitochondria. Plant Physiol. 1989 Apr;89(4):1311–1317. doi: 10.1104/pp.89.4.1311. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Guy R. D., Fogel M. L., Berry J. A. Photosynthetic Fractionation of the Stable Isotopes of Oxygen and Carbon. Plant Physiol. 1993 Jan;101(1):37–47. doi: 10.1104/pp.101.1.37. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hoefnagel M. H., Millar A. H., Wiskich J. T., Day D. A. Cytochrome and alternative respiratory pathways compete for electrons in the presence of pyruvate in soybean mitochondria. Arch Biochem Biophys. 1995 Apr 20;318(2):394–400. doi: 10.1006/abbi.1995.1245. [DOI] [PubMed] [Google Scholar]
  9. Millar A. H., Wiskich J. T., Whelan J., Day D. A. Organic acid activation of the alternative oxidase of plant mitochondria. FEBS Lett. 1993 Aug 30;329(3):259–262. doi: 10.1016/0014-5793(93)80233-k. [DOI] [PubMed] [Google Scholar]
  10. Moore A. L., Bonner W. D., Jr, Rich P. R. The determination of the proton-motive force during cyanide-insensitive respiration in plant mitochondria. Arch Biochem Biophys. 1978 Mar;186(2):298–306. doi: 10.1016/0003-9861(78)90439-3. [DOI] [PubMed] [Google Scholar]
  11. Moore A. L., Siedow J. N. The regulation and nature of the cyanide-resistant alternative oxidase of plant mitochondria. Biochim Biophys Acta. 1991 Aug 23;1059(2):121–140. doi: 10.1016/s0005-2728(05)80197-5. [DOI] [PubMed] [Google Scholar]
  12. Robinson S. A., Yakir D., Ribas-Carbo M., Giles L., Osmond C. B., Siedow J. N., Berry J. A. Measurements of the Engagement of Cyanide-Resistant Respiration in the Crassulacean Acid Metabolism Plant Kalanchoë daigremontiana with the Use of On-Line Oxygen Isotope Discrimination. Plant Physiol. 1992 Nov;100(3):1087–1091. doi: 10.1104/pp.100.3.1087. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Sakajo S., Minagawa N., Yoshimoto A. Characterization of the alternative oxidase protein in the yeast Hansenula anomala. FEBS Lett. 1993 Mar 8;318(3):310–312. doi: 10.1016/0014-5793(93)80535-3. [DOI] [PubMed] [Google Scholar]
  14. Umbach A. L., Siedow J. N. Covalent and Noncovalent Dimers of the Cyanide-Resistant Alternative Oxidase Protein in Higher Plant Mitochondria and Their Relationship to Enzyme Activity. Plant Physiol. 1993 Nov;103(3):845–854. doi: 10.1104/pp.103.3.845. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Umbach A. L., Wiskich J. T., Siedow J. N. Regulation of alternative oxidase kinetics by pyruvate and intermolecular disulfide bond redox status in soybean seedling mitochondria. FEBS Lett. 1994 Jul 11;348(2):181–184. doi: 10.1016/0014-5793(94)00600-8. [DOI] [PubMed] [Google Scholar]
  16. Weger H. G., Guy R. D., Turpin D. H. Cytochrome and alternative pathway respiration in green algae : measurements using inhibitors and o(2) discrimination. Plant Physiol. 1990 May;93(1):356–360. doi: 10.1104/pp.93.1.356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Wilson S. B. The switching of electron flux from the cyanide-insensitive oxidase to the cytochrome pathway in mung-bean (Phaseolus aureus L.) mitochondria. Biochem J. 1988 Jan 1;249(1):301–303. doi: 10.1042/bj2490301. [DOI] [PMC free article] [PubMed] [Google Scholar]

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