Skip to main content
NCBI home page
Protein Science : A Publication of the Protein Society logoLink to Protein Science : A Publication of the Protein Society
. 1992 Nov;1(11):1428–1434. doi: 10.1002/pro.5560011104

Electronic and vibrational spectroscopy of the cytochrome c:cytochrome c oxidase complexes from bovine and Paracoccus denitrificans.

S R Lynch 1, R A Copeland 1
PMCID: PMC2142114  PMID: 1338946

Abstract

The 1:1 complex between horse heart cytochrome c and bovine cytochrome c oxidase, and between yeast cytochrome c and Paracoccus denitrificans cytochrome c oxidase have been studied by a combination of second derivative absorption, circular dichroism (CD), and resonance Raman spectroscopy. The second derivative absorption and CD spectra reveal changes in the electronic transitions of cytochrome a upon complex formation. These results could reflect changes in ground state heme structure or changes in the protein environment surrounding the chromophore that affect either the ground or excited electronic states. The resonance Raman spectrum, on the other hand, reflects the heme structure in the ground electronic state only and shows no significant difference between cytochrome a vibrations in the complex or free enzyme. The only major difference between the Raman spectra of the free enzyme and complex is a broadening of the cytochrome a3 formyl band of the complex that is relieved upon complex dissociation at high ionic strength. These data suggest that the differences observed in the second derivative and CD spectra are the result of changes in the protein environment around cytochrome a that affect the electronic excited state. By analogy to other protein-chromophore systems, we suggest that the energy of the Soret pi* state of cytochrome a may be affected by (1) changes in the local dielectric, possibly brought about by movement of a charged amino acid side chain in proximity to the heme group, or (2) pi-pi interactions between the heme and aromatic amino acid residues.

Full Text

The Full Text of this article is available as a PDF (675.2 KB).

Selected References

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

  1. Brautigan D. L., Ferguson-Miller S., Margoliash E. Mitochondrial cytochrome c: preparation and activity of native and chemically modified cytochromes c. Methods Enzymol. 1978;53:128–164. doi: 10.1016/s0076-6879(78)53021-8. [DOI] [PubMed] [Google Scholar]
  2. Copeland R. A. Conformational switching at cytochrome a during steady-state turnover of cytochrome c oxidase. Proc Natl Acad Sci U S A. 1991 Aug 15;88(16):7281–7283. doi: 10.1073/pnas.88.16.7281. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Copeland R. A., Spiro T. G. Ultraviolet resonance Raman spectra of cytochrome c conformational states. Biochemistry. 1985 Aug 27;24(18):4960–4968. doi: 10.1021/bi00339a035. [DOI] [PubMed] [Google Scholar]
  4. Fowler G. J., Visschers R. W., Grief G. G., van Grondelle R., Hunter C. N. Genetically modified photosynthetic antenna complexes with blueshifted absorbance bands. Nature. 1992 Feb 27;355(6363):848–850. doi: 10.1038/355848a0. [DOI] [PubMed] [Google Scholar]
  5. Goodman G., Leigh J. S., Jr The distance between cytochromes a and a3 in the azide compound of bovine-heart cytochrome oxidase. Biochim Biophys Acta. 1987 Mar 4;890(3):360–367. doi: 10.1016/0005-2728(87)90164-2. [DOI] [PubMed] [Google Scholar]
  6. HERSKOVITS T. T., LASKOWSKI M., Jr Location of chromophoric residues in proteins by solvent perturbation. I. Tyrosyls in serum albumins. J Biol Chem. 1962 Aug;237:2481–2492. [PubMed] [Google Scholar]
  7. Hildebrandt P. G., Copeland R. A., Spiro T. G., Otlewski J., Laskowski M., Jr, Prendergast F. G. Tyrosine hydrogen-bonding and environmental effects in proteins probed by ultraviolet resonance Raman spectroscopy. Biochemistry. 1988 Jul 26;27(15):5426–5433. doi: 10.1021/bi00415a007. [DOI] [PubMed] [Google Scholar]
  8. Ishibe N., Lynch S. R., Copeland R. A. The pH dependence of cytochrome a conformation in cytochrome c oxidase. J Biol Chem. 1991 Dec 15;266(35):23916–23920. [PubMed] [Google Scholar]
  9. Ludwig B. Cytochrome c oxidase from Paracoccus denitrificans. Methods Enzymol. 1986;126:153–159. doi: 10.1016/s0076-6879(86)26017-6. [DOI] [PubMed] [Google Scholar]
  10. Lynch S. R., Sherman D., Copeland R. A. Cytochrome c binding affects the conformation of cytochrome a in cytochrome c oxidase. J Biol Chem. 1992 Jan 5;267(1):298–302. [PubMed] [Google Scholar]
  11. Michel B., Bosshard H. R. Spectroscopic analysis of the interaction between cytochrome c and cytochrome c oxidase. J Biol Chem. 1984 Aug 25;259(16):10085–10091. [PubMed] [Google Scholar]
  12. Sassaroli M., Ching Y. C., Dasgupta S., Rousseau D. L. Cytochrome c oxidase: evidence for interaction of water molecules with cytochrome a. Biochemistry. 1989 Apr 18;28(8):3128–3132. doi: 10.1021/bi00434a002. [DOI] [PubMed] [Google Scholar]
  13. Sherman D., Kotake S., Ishibe N., Copeland R. A. Resolution of the electronic transitions of cytochrome c oxidase: evidence for two conformational states of ferrous cytochrome alpha. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4265–4269. doi: 10.1073/pnas.88.10.4265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Sherman F., Stewart J. W., Parker J. H., Inhaber E., Shipman N. A., Putterman G. J., Gardisky R. L., Margoliash E. The mutational alteration of the primary structure of yeast iso-1-cytochrome c. J Biol Chem. 1968 Oct 25;243(20):5446–5456. [PubMed] [Google Scholar]
  15. Weber C., Michel B., Bosshard H. R. Spectroscopic analysis of the cytochrome c oxidase-cytochrome c complex: circular dichroism and magnetic circular dichroism measurements reveal change of cytochrome c heme geometry imposed by complex formation. Proc Natl Acad Sci U S A. 1987 Oct;84(19):6687–6691. doi: 10.1073/pnas.84.19.6687. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from Protein Science : A Publication of the Protein Society are provided here courtesy of The Protein Society

RESOURCES