title: NMR characterization of surface interactions in the cytochrome b5-cytochrome c complex.
abstract: The complex formed in solution by native and chemically modified cytochrome c with cytochrome b5 has been studied by 1H and 13C nuclear magnetic resonance spectroscopy (NMR). Contrary to predictions of recent theoretical analysis, 1H NMR spectroscopy indicates that there is no major movement of cytochrome c residue Phe82 on binding to cytochrome b5. The greater resolution provided by 13C NMR spectroscopy permits detection of small perturbations in the environments of cytochrome c residues Ile75 and Ile85 on binding with cytochrome b5, a result that is in agreement with earlier model-building experiments. As individual cytochrome c lysyl residues are resolved in the 1H NMR spectrum of N-acetimidylated cytochrome c, the interaction of this modified protein with cytochrome b5 has been studied to evaluate the number of cytochrome c lysyl residues involved in binding to cytochrome b5. The results of this experiment indicate that at least six lysyl residues are involved, two more than predicted by static model building, which indicates that cytochrome c and cytochrome b5 form two or more structurally similar 1:1 complexes in solution.
authors.affiliation: School of Chemical Sciences, University of East Anglia, Norwich, United Kingdom.
authors.forename: A M
authors.initials: AM
authors.lastname: Burch
authors.forename: S E
authors.initials: SE
authors.lastname: Rigby
authors.forename: W D
authors.initials: WD
authors.lastname: Funk
authors.forename: R T
authors.initials: RT
authors.lastname: MacGillivray
authors.forename: M R
authors.initials: MR
authors.lastname: Mauk
authors.forename: A G
authors.initials: AG
authors.lastname: Mauk
authors.forename: G R
authors.initials: GR
authors.lastname: Moore
mesh_terms: D002247:Carbon Isotopes; D003574:Cytochrome c Group / Q000378:metabolism*; D015786:Cytochromes b5 / Q000378:metabolism*; D006859:Hydrogen; D009682:Magnetic Resonance Spectroscopy / Q000379:methods; D011485:Protein Binding; D011487:Protein Conformation; D013499:Surface Properties
chemical_list: D002247:Carbon Isotopes; D003574:Cytochrome c Group; D006859:Hydrogen; D015786:Cytochromes b5