Skip to main content
Biochemical Journal logoLink to Biochemical Journal
. 1980 Nov 15;192(2):687–693. doi: 10.1042/bj1920687

The effect of complex-formation with polyanions on the redox properties of cytochrome c.

L C Petersen, R P Cox
PMCID: PMC1162385  PMID: 6165356

Abstract

1. The stable complex formed between mammalian cytochrome c and phosvitin at low ionic strength was studied by partition in an aqueous two-phase system. Oxidized cytochrome c binds to phosvitin with a higher affinity than reduced cytochrome c. The difference was equivalent to a decrease of the redox potential by 22 mV on binding. 2. Complex-formation with phosvitin strongly inhibited the reaction of cytochrome c with reagents that react as negatively charged species, such as ascorbate, dithionite, ferricyanide and tetrachlorobenzoquinol. Reaction with uncharged reagents such as NNN'N'-tetramethylphenylenediamine and the reduced form of the N-methylphenazonium ion (present as the methylsulphate) was little affected by complex-formation, whereas oxidation of the reduced cytochrome by the positively charged tris-(phenanthroline)cobalt(III) ion was greatly stimulated. 3. A similar pattern of inhibition and stimulation of reaction rates was observed when phosvitin was replaced by other macromolecular polyanions such as dextran sulphate and heparin, indicating that the results were a general property of complex-formation with polyanions. A weaker but qualitatively similar effect was observed on addition of inositol hexaphosphate and ATP. 4. It is suggested that the effects of complex-formation with polyanions on the reactivity of cytochrome c with redox reagents are mainly the result of replacing the positive charge on the free cytochrome by a net negative charge. Any steric effects on polyanion binding are small in comparison with such electrostatic effects.

Full text

PDF
687

Selected References

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

  1. Al-Ayash A. I., Wilson M. T. The mechanism of reduction of single-site redox proteins by ascorbic acid. Biochem J. 1979 Feb 1;177(2):641–648. doi: 10.1042/bj1770641. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Allerton S. E., Perlmann G. E. Chemical characterization of the phosphoprotein phosvitin. J Biol Chem. 1965 Oct;240(10):3892–3898. [PubMed] [Google Scholar]
  3. Barlow G. H., Margoliash E. Electrophoretic behavior of mammalian-type cytochromes c. J Biol Chem. 1966 Apr 10;241(7):1473–1477. [PubMed] [Google Scholar]
  4. Cassatt J. C., Marini C. P. The kinetics of oxidation of reduced cytochrome c by ferricyanide derivatives. Biochemistry. 1974 Dec 17;13(26):5323–5328. doi: 10.1021/bi00723a011. [DOI] [PubMed] [Google Scholar]
  5. Giancotti V., Quadrifoglio F., Crescenzi V. Polyelectrolyte behaviour of phosvitin. Spectroscopic, microcalorimetric and acridine-orange-binding data. Eur J Biochem. 1973 May;35(1):78–86. doi: 10.1111/j.1432-1033.1973.tb02812.x. [DOI] [PubMed] [Google Scholar]
  6. Goldkorn T., Schejter A. Electrostatic effects on the kinetics of oxidation-reduction reactions of c-type cytochromes. J Biol Chem. 1979 Dec 25;254(24):12562–12566. [PubMed] [Google Scholar]
  7. Goldkorn T., Schejter A. Kinetic and spectroscopic evidence for different forms of ferric cytochrome c at very low ionic strength and neutral pH. FEBS Lett. 1977 Mar 15;75(1):44–46. doi: 10.1016/0014-5793(77)80049-5. [DOI] [PubMed] [Google Scholar]
  8. Kihara H., Nakatani H., Hiromi K., Hon-Nami K., Oshima T. Kinetic studies on redox reactions of hemoproteins. II. Reduction of thermoresistant cytochrome c-552 and horse heart cytochrome c by ascorbic acid. J Biochem. 1978 Jan;83(1):243–248. doi: 10.1093/oxfordjournals.jbchem.a131897. [DOI] [PubMed] [Google Scholar]
  9. MINNAERT K. The kinetics of cytochrome c oxidase. II. The system: reducing agent-cytochrome c-cytochrome oxidase-oxygen. Biochim Biophys Acta. 1961 Nov 25;54:26–41. doi: 10.1016/0006-3002(61)90934-9. [DOI] [PubMed] [Google Scholar]
  10. Margalit R., Schejter A. Cytochrome c: a thermodynamic study of relationships among oxidation state, ion-binding and structural parameters. 2. Ion-binding linked to oxidation state. Eur J Biochem. 1973 Feb 1;32(3):500–505. doi: 10.1111/j.1432-1033.1973.tb02634.x. [DOI] [PubMed] [Google Scholar]
  11. Margalit R., Schejter A. Cytochrome c: a thermodynamic study of the relationships among oxidation state, ion-binding and structural parameters. 1. The effects of temperature, pH and electrostatic media on the standard redox potential of cytochrome c. Eur J Biochem. 1973 Feb 1;32(3):492–499. doi: 10.1111/j.1432-1033.1973.tb02633.x. [DOI] [PubMed] [Google Scholar]
  12. Margoliash E., Barlow G. H., Byers V. Differential binding properties of cytochrome c: possible relevance for mitochondrial ion transport. Nature. 1970 Nov 21;228(5273):723–726. doi: 10.1038/228723a0. [DOI] [PubMed] [Google Scholar]
  13. McArdle J. V., Gray H. B., Creutz C., Sutin N. Kinetic studies of the oxidation of ferrocytochrome c from horse heart and Candida krusei by tris(1,10-phenanthroline)cobalt(3). J Am Chem Soc. 1974 Sep 4;96(18):5737–5741. doi: 10.1021/ja00825a009. [DOI] [PubMed] [Google Scholar]
  14. Miller W. G., Cusanovich M. A. Electron transport by C-type cytochromes. I. The reaction of horse heart cytochrome c with anionic reductants. Biophys Struct Mech. 1975 Feb 19;1(2):97–111. [PubMed] [Google Scholar]
  15. Mochan E., Nicholls P. Cytochrome c reactivity in its complexes with mammalian cytochrome c oxidase and yeast peroxidase. Biochim Biophys Acta. 1972 May 25;267(2):309–319. doi: 10.1016/0005-2728(72)90119-3. [DOI] [PubMed] [Google Scholar]
  16. Morton R. A., Overnell J., Harbury H. A. Electron transfer between cytochromes c from horse and Pseudomonas. J Biol Chem. 1970 Sep 25;245(18):4653–4657. [PubMed] [Google Scholar]
  17. Nicholls P. Cytochrome c binding to enzymes and membranes. Biochim Biophys Acta. 1974 Dec 30;346(3-4):261–310. doi: 10.1016/0304-4173(74)90003-2. [DOI] [PubMed] [Google Scholar]
  18. Peterman B. F., Morton R. A. The effect of binding ions on the oxidation of horse heart ferrocytochrome c. Can J Biochem. 1979 May;57(5):372–377. doi: 10.1139/o79-047. [DOI] [PubMed] [Google Scholar]
  19. Petersen L. C., Cox R. P. On the mechanism of the cytochrome c oxidase reaction. Bound Fe(III) cytochrome c as an intermediate in the reductive half reaction. Eur J Biochem. 1980 Apr;105(2):321–327. doi: 10.1111/j.1432-1033.1980.tb04504.x. [DOI] [PubMed] [Google Scholar]
  20. Petersen L. C. Cytochrome c--cytochrome aa3 complex formation at low ionic strength studied by aqueous two-phase partition. FEBS Lett. 1978 Oct 1;94(1):105–108. doi: 10.1016/0014-5793(78)80916-8. [DOI] [PubMed] [Google Scholar]
  21. Rich P. R., Bendall D. S. A mechanism for the reduction of cytochromes by quinols in solution and its relevance to biological electron transfer reactions. FEBS Lett. 1979 Sep 15;105(2):189–194. doi: 10.1016/0014-5793(79)80608-0. [DOI] [PubMed] [Google Scholar]
  22. Rieder R., Bosshard H. R. The cytochrome c oxidase binding site on cytochrome c. Differential chemical modification of lysine residues in free and oxidase-bound cytochrome c. J Biol Chem. 1978 Sep 10;253(17):6045–6053. [PubMed] [Google Scholar]
  23. Schejter A., Margalit R. The redox potential of cytochrome c: Ion binding and oxidation state as linked functions. FEBS Lett. 1970 Oct 5;10(3):179–181. doi: 10.1016/0014-5793(70)80447-1. [DOI] [PubMed] [Google Scholar]
  24. Smith L., Davies H. C., Nava M. E. Studies of the kinetics of oxidation of cytochrome c by cytochrome c oxidase: comparison of spectrophotometric and polarographic assays. Biochemistry. 1979 Jul 10;18(14):3140–3146. doi: 10.1021/bi00581a035. [DOI] [PubMed] [Google Scholar]
  25. Taborsky G. Interaction of cytochrome c and the phosphoprotein phosvitin. Formation of a complex with an intact 695-mmicron absorption band. Biochemistry. 1970 Sep 15;9(19):3768–3774. doi: 10.1021/bi00821a016. [DOI] [PubMed] [Google Scholar]
  26. Vanderkooi J., Erecińska M., Chance B. Cytochrome c interaction with membranes. I. Use of a fluorescent chromophore in the study of cytochrome c interaction with artificial and mitochondrial membranes. Arch Biochem Biophys. 1973 Jan;154(1):219–229. doi: 10.1016/0003-9861(73)90052-0. [DOI] [PubMed] [Google Scholar]
  27. Weiss H., Juchs B. Isolation of a multiprotein complex containing cytochrome b and c1 from Neurospora crassa mitochondria by affinity chromatography on immobilized cytochrome c. Difference in the binding between ferricytochrome c and ferrocytochrome c to the multiprotein complex. Eur J Biochem. 1978 Jul 17;88(1):17–28. doi: 10.1111/j.1432-1033.1978.tb12418.x. [DOI] [PubMed] [Google Scholar]
  28. YAMAZAKI I. The reduction of cytochrome c by enzyme-generated ascorbic free radical. J Biol Chem. 1962 Jan;237:224–229. [PubMed] [Google Scholar]
  29. Yoshimura T., Matsushima A., Aki K. Interaction of cytochrome c with the phosphorprotein phosvitin. Biochim Biophys Acta. 1979 Dec 14;581(2):316–324. doi: 10.1016/0005-2795(79)90251-4. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES