Dynamic docking and electron transfer between myoglobin and cytochrome b5

Zhao Xun Liang, Min Jiang, Qing Ning, Brian M. Hoffman*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

40 Scopus citations

Abstract

The interaction of trypsin-digested bovine cytochrome b5 (cyt b5) with horse heart myoglobin (Mb) and the interprotein electron transfer (ET) between these redox partners have been studied to gain better understanding of ET processes between weakly bound protein partners. The bimolecular rate constant (k2) for photo-induced ET between zinc-substituted Mb (ZnMb) and cyt b5 decreases with increasing ionic strength, consistent with the predominantly electrostatic character of this complex. The formation of a protein-protein complex has been confirmed and the binding affinities of metMb and ZnMb for cyt b5 have been measured by two techniques: 1H NMR titrations at pH 6.0 give binding constants of Ka≈(1.0±0.1)×103 M-1 for metMb and Ka≈(0.75±0.1)×103 M-1 for ZnMb; isothermal calorimetry gives Ka≈(0.35±0.1)×103 M-1 for ZnMb. Brownian dynamic (BD) simulations show that cyt b5 binds over a broad surface of Mb that includes its heme edge. The experimental results are described in terms of a dynamic docking model which proposes that Mb binds cyt b5 in a large ensemble of protein binding conformations, not one or a few dominant ones, but that only a small subset are ET reactive. Aided by the BD simulations, this model explains why k2 decreases with increasing pH: increasing pH not only weakens the binding affinity but also reduces the number of binding conformations with high ET reactivity.

Original languageEnglish (US)
Pages (from-to)580-588
Number of pages9
JournalJournal of Biological Inorganic Chemistry
Volume7
Issue number6
DOIs
StatePublished - 2002

Keywords

  • Brownian dynamic simulation
  • Cytochrome b
  • Electron transfer
  • Myoglobin
  • Protein interaction

ASJC Scopus subject areas

  • Biochemistry
  • Inorganic Chemistry

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