title: The regulation of surface charged residues on the properties of cytochrome b5.
abstract: To understand the roles of negatively surface charged residues, the cytochrome b5 (Cyt b5) E48A/D60A mutant was constructed. UV-visible and CD spectra confirmed that the mutation did not cause overall structural changes of the protein. The mutant presents an unexpected high stability toward the thermal and denaturant compared with the wild type Cyt b5, which shows that these surface charged residues can influence the interactions between the heme b group and the polypeptide chain. Functional properties were clarified through the electron transfer reactions between Cyt b5 and Cyt c. The driving force of the electron transfer reactions is conservative. Although the association constant of Cyt b5 E48A/D60A with Cyt c is much lower than that of the wild type Cyt b5, their electron transfer rate constants do not differ significantly. The results show that these surface charged residues play important roles in regulating both the stability and functional properties of Cyt b5.
authors.affiliation: Chemical Biology Laboratory, Chemistry Department of Fudan University, Shanghai, PR China.
authors.forename: Y H
authors.initials: YH
authors.lastname: Wang
authors.forename: Y
authors.initials: Y
authors.lastname: Ren
authors.forename: W H
authors.initials: WH
authors.lastname: Wang
authors.forename: Y
authors.initials: Y
authors.lastname: Xie
authors.forename: Z X
authors.initials: ZX
authors.lastname: Huang
mesh_terms: D001483:Base Sequence; D002942:Circular Dichroism; D015786:Cytochromes b5 / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D017931:DNA Primers; D016297:Mutagenesis, Site-Directed; D021241:Spectrometry, Mass, Electrospray Ionization; D013056:Spectrophotometry, Ultraviolet; D013499:Surface Properties
chemical_list: D017931:DNA Primers; D015786:Cytochromes b5