Abstract
The oxidative and phosphorylative properties of mitochondria isolated from Neurospora crassa were investigated as a function of growth stage. The rates of oxidation of exogenous NADH and NADPH varied independently of each other, thus ruling out the existence of only one unspecific dehydrogenase. Two different pathways were involved in the oxidation of NAD-linked substrates, as indicated by changes in the rate of oxygen uptake, the sensitivity to rotenone, and the efficiency of phosphorylation. One pathway was sensitive to rotenone and involved three energy-coupling sites, whereas the other was resistant to rotenone and bypassed complex I. Our results indicated that the activity of complex I of the respiratory chain increased markedly in the late exponential phase of growth, remained high in the stationary phase, and then decreased when conidiae were formed. In contrast, the activity of the rotenone-resistant bypass was maximal in the early exponential phase. With malate (plus glutamate) as a substrate, the sensitivity to rotenone and the ADP/O ratios were always lower than those observed with other NAD-linked substrates, suggesting a possible cooperation between malate dehydrogenase and the rotenone-resistant pathway. The rate of oxygen uptake measured in the presence of rotenone was significantly increased by the addition of exogenous NAD+, suggesting that added NAD+ could interact with the rotenone-resistant bypass.
Full text
PDFSelected References
These references are in PubMed. This may not be the complete list of references from this article.
- Brunton C. J., Palmer J. M. Pathways for the oxidation of malate and reduced pyridine nucleotide by wheat mitochondria. Eur J Biochem. 1973 Nov 1;39(1):283–291. doi: 10.1111/j.1432-1033.1973.tb03125.x. [DOI] [PubMed] [Google Scholar]
- CHANCE B., WILLIAMS G. R. Respiratory enzymes in oxidative phosphorylation. I. Kinetics of oxygen utilization. J Biol Chem. 1955 Nov;217(1):383–393. [PubMed] [Google Scholar]
- Coleman J. O., Palmer J. M. The oxidation of malate by isolated plant mitochondria. Eur J Biochem. 1972 Apr 24;26(4):499–509. doi: 10.1111/j.1432-1033.1972.tb01792.x. [DOI] [PubMed] [Google Scholar]
- Day D. A., Wiskich J. T. The Effect of Exogenous Nicotinamide Adenine Dinucleotide on the Oxidation of Nicotinamide Adenine Dinucleotide-linked Substrates by Isolated Plant Mitochondria. Plant Physiol. 1974 Sep;54(3):360–363. doi: 10.1104/pp.54.3.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Day D. A., Wiskich J. T. The oxidation of malate and exogenous reduced nicotinamide adenine dinucleotide by isolated plant mitochondria. Plant Physiol. 1974 Jan;53(1):104–109. doi: 10.1104/pp.53.1.104. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Douce R., Mannella C. A., Bonner W. D., Jr The external NADH dehydrogenases of intact plant mitochondria. Biochim Biophys Acta. 1973 Jan 18;292(1):105–116. doi: 10.1016/0005-2728(73)90255-7. [DOI] [PubMed] [Google Scholar]
- Grossman S., Cobley J. G., Singer T. P. Reduced nicotinamide adenine dinucleotide dehydrogenase, piericidin sensitivity, and site 1 phosphorylation in different growth phases of Candida utilis. J Biol Chem. 1974 Jun 25;249(12):3819–3826. [PubMed] [Google Scholar]
- Hall D. O., Greenawalt J. W. The preparation and biochemical properties of mitochondria from Neurospora crassa. J Gen Microbiol. 1967 Sep;48(3):419–430. doi: 10.1099/00221287-48-3-419. [DOI] [PubMed] [Google Scholar]
- Katz R., Kilpatrick L., Chance B. Acquisition and loss of rotenone sensitivity in Torulopsis utilis. Eur J Biochem. 1971 Aug 16;21(3):301–307. doi: 10.1111/j.1432-1033.1971.tb01470.x. [DOI] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lambowitz A. M., Smith E. W., Slayman C. W. Electron transport in Neurospora mitochondria. Studies on wild type and poky. J Biol Chem. 1972 Aug 10;247(15):4850–4858. [PubMed] [Google Scholar]
- Lambowitz A. M., Smith E. W., Slayman C. W. Oxidative phosphorylation in Neurospora mitochondria. Studies on wild type, poky, and chloramphenicol-induced wild type. J Biol Chem. 1972 Aug 10;247(15):4859–4865. [PubMed] [Google Scholar]
- Munkres K. D., Woodward D. O. Interaction of Neurospora mitochondrial structural protein with other proteins and coenzyme nucleotides. Biochim Biophys Acta. 1967 Jan 18;133(1):143–150. doi: 10.1016/0005-2795(67)90046-3. [DOI] [PubMed] [Google Scholar]
- Onishi T. Mechanism of electron transport and energy conservation in the site I region of the respiratory chain. Biochim Biophys Acta. 1973 Dec 7;301(2):105–128. [PubMed] [Google Scholar]
- Riesz P., White F. H., Jr Determination of free radicals in gamma irradiated proteins. Nature. 1967 Dec 23;216(5121):1208–1210. doi: 10.1038/2161208b0. [DOI] [PubMed] [Google Scholar]
- Rustin P., Moreau F., Lance C. Malate Oxidation in Plant Mitochondria via Malic Enzyme and the Cyanide-insensitive Electron Transport Pathway. Plant Physiol. 1980 Sep;66(3):457–462. doi: 10.1104/pp.66.3.457. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tobin A., Djerdjour B., Journet E., Neuburger M., Douce R. Effect of NAD on Malate Oxidation in Intact Plant Mitochondria. Plant Physiol. 1980 Aug;66(2):225–229. doi: 10.1104/pp.66.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Weiss H., von Jagow G., Klingenberg M., Bücher T. Characterization of Neurospora crassa mitochondria prepared with a grind-mill. Eur J Biochem. 1970 May 1;14(1):75–82. doi: 10.1111/j.1432-1033.1970.tb00263.x. [DOI] [PubMed] [Google Scholar]
- von Jagow G., Klingenberg M. Pathways of hydrogen in mitochondria of Saccharomyces carlsbergensis. Eur J Biochem. 1970 Feb;12(3):583–592. doi: 10.1111/j.1432-1033.1970.tb00890.x. [DOI] [PubMed] [Google Scholar]