Revelation of ternary complexes between redox partners in cytochrome P450-containing monooxygenase systems by the optical biosensor method

Yuri D. Ivanov, Irina P. Kanaeva, Irina I. Karuzina, Sergey A. Usanov, Gaston Hui Bon Hoa, Stephen G. Sligar, Alexander I. Archakov

Research output: Contribution to journalArticle

Abstract

Formation of binary and ternary complexes in the water-soluble cytochrome P450cam (P450cam)-containing as well as in the membrane P4502B4(2B4)- and the mixed P450scc-containing monooxygenase systems was investigated in real time by the 'resonant mirror' optical biosensor method. It was shown that the inter-protein electron transfer occurs not only during complex formation but also upon random collision - as was the case with the d-Fp/d-b5 pair (2B4 system). Binary complexes may be either facilitative to electron transfer (electron-transfer complexes) or prohibitive to it (non-productive complexes). Although the binary PdR/Pd and P450cam/Pd complex formation (within the P450cam-system) as well as the binary AdR/Ad and P450scc/Ad complex formation (within the P450scc-system) does occur, the lifetimes of these complexes formed are several orders of magnitude higher than the time required for realization of a complete hydroxylation cycle. At the same time, the lifetimes of the ternary PdR/Pd/P450cam and AdR/Ad/P450scc complexes are sufficient to permit the realization of a complete hydroxylation cycle in either of these systems. For the membrane P450 2B4 system, the formation of both the binary (Fp/2B4 and 2B4/b5) and ternary (Fp/2B4/b5) complexes was registered. The lifetimes of the binary Fp/2B4 and the ternary Fp/2B4/b5 complexes are sufficient for realization of a complete hydroxylation cycle in each of them.

Original languageEnglish (US)
Pages (from-to)175-184
Number of pages10
JournalJournal of Inorganic Biochemistry
Volume87
Issue number4
DOIs
StatePublished - Dec 15 2001

Keywords

  • Cytochrome P450
  • Optical biosensor
  • Ternary complexes

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

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