title: Identification of the quinone cofactor in a lysyl oxidase from Pichia pastoris.
abstract: A copper amine oxidase from Pichia pastoris is the only known non-mammalian lysyl oxidase [Tur, S.S. and Lerch, K. (1988) FEBS Lett. 238, 74-76]. Recently, the cofactor in mammalian lysyl oxidase has been identified as a novel lysine tyrosylquinone moiety [Wang, S.X., Mure, M., Medzihradszky, K.F., Burlingame, A.L., Brown, D.E., Dooley, D.M., Smith, A.J., Kagan, H.M. and Klinman, J.P. (1996) Science 273, 1078-1084]. In order to identify the cofactor in P. pastoris lysyl oxidase, we have isolated the phenylhydrazone-derivative of the active-site peptide. This peptide has the active-site sequence conserved among topa quinone containing amine oxidases. The resonance Raman spectra of the phenylhydrazone derivatives of the enzyme, active-site peptide, and a topa quinone model compound are essentially identical. Collectively, these results establish that P. pastoris lysyl oxidase is a topa quinone enzyme.
authors.affiliation: Department of Chemistry, University of California, Berkeley 94720, USA.
authors.forename: J E
authors.initials: JE
authors.lastname: Dove
authors.forename: A J
authors.initials: AJ
authors.lastname: Smith
authors.forename: J
authors.initials: J
authors.lastname: Kuchar
authors.forename: D E
authors.initials: DE
authors.lastname: Brown
authors.forename: D M
authors.initials: DM
authors.lastname: Dooley
authors.forename: J P
authors.initials: JP
authors.lastname: Klinman
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002851:Chromatography, High Pressure Liquid; D004295:Dihydroxyphenylalanine / Q000031:analogs & derivatives* / Q000032:analysis; D006835:Hydrazones; D010843:Pichia / Q000201:enzymology*; D008249:Protein-Lysine 6-Oxidase / Q000737:chemistry* / Q000302:isolation & purification; D013059:Spectrum Analysis, Raman
chemical_list: D006835:Hydrazones; C012483:phenylhydrazone; D004295:Dihydroxyphenylalanine; C067626:6-hydroxydopa quinone; D008249:Protein-Lysine 6-Oxidase