title: NMR and kinetic characterization of the interaction between French bean plastocyanin and horse cytochrome c.
abstract: French bean plastocyanin is shown by stopped-flow kinetics to oxidize horse cytochrome c with k (298 K, I = 0.10 M) = 5.1 X 10(6) M-1 X s-1. The activation parameters demonstrate a satisfactory isokinetic correlation with those previously reported for plastocyanin-cytochrome f reactions. NMR line broadening and shifts of the hyperfine shifted resonances of cytochrome c(III) reveal that strong 1:1 complexes are formed with plastocyanin. The negative patch of plastocyanin and the heme edge region of cytochrome c are shown to be the interacting sites by the hyperfine shift perturbations and competitive binding experiments with Gd3+, which associates selectively with the negative patch of plastocyanin. Complexation of plastocyanin and cytochrome c causes a small change in the heme electronic structure, but there is no NMR or optical evidence for significant conformational changes at either metal center. The rate of the reverse electron-transfer reaction within the plastocyanin-cytochrome c complex has been directly measured by NMR line broadening (krev (298 K) = 87 s-1). A rate for the forward intracomplex electron-transfer reaction (kf (298 K) = 4.8 X 10(3) s-1) has been calculated from krev and the optically measured equilibrium constant.
authors.forename: G C
authors.initials: GC
authors.lastname: King
authors.forename: R A
authors.initials: RA
authors.lastname: Binstead
authors.forename: P E
authors.initials: PE
authors.lastname: Wright
mesh_terms: D000818:Animals; D001665:Binding Sites; D003300:Copper; D003574:Cytochrome c Group / Q000378:metabolism*; D005296:Ferrous Compounds; D006736:Horses; D007700:Kinetics; D009682:Magnetic Resonance Spectroscopy; D010940:Plant Proteins / Q000378:metabolism*; D010944:Plants; D010970:Plastocyanin / Q000378:metabolism*; D013696:Temperature
chemical_list: D003574:Cytochrome c Group; D005296:Ferrous Compounds; D010940:Plant Proteins; D003300:Copper; D010970:Plastocyanin