title: The cytochrome P450 gene family CYP157 does not contain EXXR in the K-helix reducing the absolute conserved P450 residues to a single cysteine.
abstract: In this work, we have spectroscopically characterised CYP157C1 from Streptomyces coelicolor A3(2) which has the motif E(297)QSLW(301) rather than the invariant EXXR motif in the P450 K-helix. Site-directed mutagenesis of native E(297)QSLW(301) in CYP157C1 to E(297)ESLR(301) or E(297)QSRW(301) both containing standard EXXR motifs produced cytochrome P420 proteins thought to be inactive forms of P450 even though wild type CYP157C1 has the spectral properties of a normal P450. These results indicate that the EXXR motif is not required in all CYP tertiary architectures and only a single cysteine residue, which coordinates as the fifth thiolate ligand to the P450 haem iron, is invariant in all CYPs structures.
authors.affiliation: Department of Cell and Developmental Biology, University of Illinois, 1201 W Gregory Dr, Urbana, IL 61801, USA.
authors.forename: Sanjeewa
authors.initials: S
authors.lastname: Rupasinghe
authors.forename: Mary A
authors.initials: MA
authors.lastname: Schuler
authors.forename: Norio
authors.initials: N
authors.lastname: Kagawa
authors.forename: Hang
authors.initials: H
authors.lastname: Yuan
authors.forename: Li
authors.initials: L
authors.lastname: Lei
authors.forename: Bin
authors.initials: B
authors.lastname: Zhao
authors.forename: Steven L
authors.initials: SL
authors.lastname: Kelly
authors.forename: Michael R
authors.initials: MR
authors.lastname: Waterman
authors.forename: David C
authors.initials: DC
authors.lastname: Lamb
mesh_terms: D020816:Amino Acid Motifs / Q000235:genetics; D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics; D003545:Cysteine / Q000737:chemistry / Q000235:genetics; D003577:Cytochrome P-450 Enzyme System / Q000737:chemistry* / Q000235:genetics; D006418:Heme / Q000737:chemistry / Q000235:genetics; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed / Q000379:methods; D020836:Protein Structure, Quaternary / Q000235:genetics; D013056:Spectrophotometry, Ultraviolet / Q000379:methods; D048372:Streptomyces coelicolor / Q000201:enzymology* / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D006418:Heme; D003577:Cytochrome P-450 Enzyme System; D003545:Cysteine