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. 1986 Sep;82(1):1–6. doi: 10.1104/pp.82.1.1

Characterization and Solubilization of the Alternative Oxidase of Sauromatum guttatum Mitochondria 1

Thomas E Elthon 1, Lee McIntosh 1
PMCID: PMC1056056  PMID: 16664973

Abstract

The alternative oxidase activity of Sauromatum guttatum spadix mitochondria has been investigated as to its developmental expression and tissue localization. Mitochondria rich in alternative oxidase activity were found in a yellow cortex tissue present to varying degrees within the appendix, male floral, and sterile floral regions of the spadix. During a 5-day period just prior to anthesis, the alternative oxidase activity present in the appendix region was found to increase over 10-fold. On the following day when the appendix region becomes thermogenic, cytochrome oxidase activity was found to decrease by 92%, effectively forcing electron flow through the alternative oxidase. A procedure for efficient solubilization of the alternative oxidase from appendix region mitochondria was developed. The alternative oxidase thus solubilized was sensitive to heat inactivation and trypsin digestion. The activity showed inhibition characteristics expected of the alternative oxidase in that it was sensitive to salicylhydroxamic acid and 5-n-undecyl-6-hydroxy-4,7-dioxobenzothiazole, but relatively insensitive to KCN and antimycin A. Essential sulfhydryl group(s) were indicated by reversible inhibition by p-chloromercuribenzoic acid. The solubilized alternative oxidase was most active in the detergent N,N-bis-(3-d-glucoamidopropyl)-deoxycholamide and had a pH optimum of 6.8.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. BRUINSMA J. A comment on the spectrophotometric determination of chlorophyll. Biochim Biophys Acta. 1961 Sep 30;52:576–578. doi: 10.1016/0006-3002(61)90418-8. [DOI] [PubMed] [Google Scholar]
  2. Bahr J. T., Bonner W. D., Jr Cyanide-insensitive respiration. I. The steady states of skunk cabbage spadix and bean hypocotyl mitochondria. J Biol Chem. 1973 May 25;248(10):3441–3445. [PubMed] [Google Scholar]
  3. Bower T. G. The visual world of infants. Sci Am. 1966 Dec;215(6):80–passim. doi: 10.1038/scientificamerican1266-80. [DOI] [PubMed] [Google Scholar]
  4. Douce R., Christensen E. L., Bonner W. D., Jr Preparation of intaintact plant mitochondria. Biochim Biophys Acta. 1972 Aug 17;275(2):148–160. doi: 10.1016/0005-2728(72)90035-7. [DOI] [PubMed] [Google Scholar]
  5. Elthon T. E., Stewart C. R., McCoy C. A., Bonner W. D. Alternative Respiratory Path Capacity in Plant Mitochondria: Effect of Growth Temperature, the Electrochemical Gradient, and Assay pH. Plant Physiol. 1986 Feb;80(2):378–383. doi: 10.1104/pp.80.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Elthon T. E., Stewart C. R. Proline Oxidation in Corn Mitochondria : Involvement of NAD, Relationship to Ornithine Metabolism, and Sidedness on the Inner Membrane. Plant Physiol. 1982 Aug;70(2):567–572. doi: 10.1104/pp.70.2.567. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Hjelmeland L. M., Chrambach A. Solubilization of functional membrane proteins. Methods Enzymol. 1984;104:305–318. doi: 10.1016/s0076-6879(84)04097-0. [DOI] [PubMed] [Google Scholar]
  8. Huq S., Palmer J. M. Isolation of a cyanide-resistant duroquinol oxidase from Arum maculatum mitochondria. FEBS Lett. 1978 Nov 15;95(2):217–220. doi: 10.1016/0014-5793(78)80997-1. [DOI] [PubMed] [Google Scholar]
  9. Kay C. J., Palmer J. M. Solubilization of the alternative oxidase of cuckoo-pint (Arum maculatum) mitochondria. Stimulation by high concentrations of ions and effects of specific inhibitors. Biochem J. 1985 Jun 1;228(2):309–318. doi: 10.1042/bj2280309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. 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]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. Moore A. L., Bonner W. D. Measurements of membrane potentials in plant mitochondria with the safranine method. Plant Physiol. 1982 Nov;70(5):1271–1276. doi: 10.1104/pp.70.5.1271. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Schwitzguebel J. P., Siegenthaler P. A. Purification of peroxisomes and mitochondria from spinach leaf by percoll gradient centrifugation. Plant Physiol. 1984 Jul;75(3):670–674. doi: 10.1104/pp.75.3.670. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Storey B. T., Bahr J. T. The respiratory chain of plant mitochondria. II. Oxidative phosphorylation in skunk cabbage mitochondria. Plant Physiol. 1969 Jan;44(1):126–134. doi: 10.1104/pp.44.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. van Renswoude J., Kempf C. Purification of integral membrane proteins. Methods Enzymol. 1984;104:329–339. doi: 10.1016/s0076-6879(84)04099-4. [DOI] [PubMed] [Google Scholar]

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