title: Structural studies of imidazole-cytochrome c: resonance assignments and structural comparison with cytochrome c.
abstract: Two-dimensional nuclear magnetic resonance spectroscopy (2D NMR) was used to obtain extensive proton resonance assignments of Im-cyt c complex which is a possible analog of a late folding intermediate of cytochrome c. Assignments were made nearly completely for the main-chain and the side-chain protons (all except Gly29). As starting points for the assignment of the Im-cyt c, a limited set of protons was initially assigned by use of 2D NMR magnetization transfer methods to correlated resonances in the Im-cyt c with assigned resonances in the native cyt c. The subsequent search focused on recognition of main-chain NOE connectivity patterns, with use of previously assigned residues to place NOE-connected segments within the amino acid sequence. The observed patterns of main-chain NOEs provided some structural information and suggested potentially significant differences between Im-cyt c and the native cyt c. Differences in NOEs involving side-chain protons were reported and analyzed. There was evidence for conformational changes induced by the breakage of Fe-S bond. It was concluded that the Im-cyt c had undergone a rearrangement of several regions forming the heme pocket of the protein. The structural understanding of these effects of the mutation may be essential to elucidate the changes in function and kinetic mechanism of cyt c folding.
authors.affiliation: State Key Laboratory of Coordination Chemistry, Nanjing University, China.
authors.forename: G
authors.initials: G
authors.lastname: Liu
authors.forename: W
authors.initials: W
authors.lastname: Shao
authors.forename: X
authors.initials: X
authors.lastname: Huang
authors.forename: H
authors.initials: H
authors.lastname: Wu
authors.forename: W
authors.initials: W
authors.lastname: Tang
mesh_terms: D000818:Animals; D003574:Cytochrome c Group / Q000737:chemistry*; D006736:Horses; D007093:Imidazoles / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D017433:Protein Structure, Secondary
chemical_list: D003574:Cytochrome c Group; D007093:Imidazoles