title: The role of cytochrome b5 structural domains in interaction with cytochromes P450.
abstract: To understand the role of the structural elements of cytochrome b5 in its interaction with cytochrome P450 and the catalysis performed by this heme protein, we carried out comparative structural and functional analysis of the two major mammalian forms of membrane-bound cytochrome b5 - microsomal and mitochondrial, designed chimeric forms of the heme proteins in which the hydrophilic domain of one heme protein is replaced by the hydrophilic domain of another one, and investigated the effect of the highly purified native and chimeric heme proteins on the enzymatic activity of recombinant cytochromes P4503A4 and P45017A1 (CYP3A4 and CYP17A1). We show that the presence of a hydrophobic domain in the structure of cytochrome b5 is necessary for its effective interaction with its redox partners, while the nature of the hydrophobic domain has no significant effect on the ability of cytochrome b5 to stimulate the activity of cytochrome P450-catalyzed reactions. Thus, the functional properties of cytochrome b5 are mainly determined by the structure of the heme-binding domain.
authors: Institute of Bioorganic Chemistry, Academy of Sciences of Belarus, Minsk, 220141, Belarus. gvserg@iboch.bas-net.by.
authors: G V
authors: GV
authors: Sergeev
authors: A A
authors: AA
authors: Gilep
authors: S A
authors: SA
authors: Usanov
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D055162:Biocatalysis; D051544:Cytochrome P-450 CYP3A / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015786:Cytochromes b5 / Q000737:chemistry / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D007700:Kinetics; D008861:Microsomes / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism; D008969:Molecular Sequence Data; D051381:Rats; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics; D013254:Steroid 17-alpha-Hydroxylase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D011994:Recombinant Proteins; D015786:Cytochromes b5; D003577:Cytochrome P-450 Enzyme System; D051544:Cytochrome P-450 CYP3A; C582555:CYP17A1 protein, rat; D013254:Steroid 17-alpha-Hydroxylase