Mapping the cytochrome c553 interacting site using 1H and 15N NMR

FEBS Lett. 1999 Oct 22;460(1):77-80. doi: 10.1016/s0014-5793(99)01299-5.

Abstract

Cytochrome c553 is the electron transfer partner of formate dehydrogenase and of [Fel-hydrogenase, two metalloenzymes essential in the metabolism of sulfate reducing bacteria. These two enzymes contain a 'ferredoxin-like' domain which presents 30% identity with Desulfovibrio desulfuricans Norway ferredoxin 1. This was chosen as a model for the 'ferredoxin-like' domain involved in the electron transfer reaction with cytochrome c553. ID NMR titration of complex formation gave us the stoichiometry (1:1) and the dissociation constant of the complex (Kd approximately 3x10(-6) M). 2D heteronuclear NMR experiments were performed to analyze the 1H and 15N chemical shift variations that are induced by the protein-protein recognition. This is the first mapping of the interaction site on a c-type cytochrome, using heteronuclear NMR.

MeSH terms

  • Bacterial Proteins / chemistry
  • Binding Sites
  • Cytochrome c Group / chemistry*
  • Desulfovibrio / enzymology*
  • Ferredoxins / chemistry
  • Formate Dehydrogenases / chemistry
  • Hydrogenase / chemistry
  • Magnetic Resonance Spectroscopy
  • Models, Molecular

Substances

  • Bacterial Proteins
  • Cytochrome c Group
  • Ferredoxins
  • cytochrome c553
  • Hydrogenase
  • Formate Dehydrogenases
  • Formate dehydrogenase (cytochrome)