title: X-ray structure of a putative reaction intermediate of 5-aminolaevulinic acid dehydratase.
abstract: The X-ray structure of yeast 5-aminolaevulinic acid dehydratase, in which the catalytic site of the enzyme is complexed with a putative cyclic intermediate composed of both substrate moieties, has been solved at 0.16 nm (1.6 A) resolution. The cyclic intermediate is bound covalently to Lys(263) with the amino group of the aminomethyl side chain ligated to the active-site zinc ion in a position normally occupied by a catalytic hydroxide ion. The cyclic intermediate is catalytically competent, as shown by its turnover in the presence of added substrate to form porphobilinogen. The findings, combined with those of previous studies, are consistent with a catalytic mechanism in which the C-C bond linking both substrates in the intermediate is formed before the C-N bond.
authors.affiliation: Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton, SO16 7PX, UK.
authors.forename: Peter T
authors.initials: PT
authors.lastname: Erskine
authors.forename: Leighton
authors.initials: L
authors.lastname: Coates
authors.forename: Danica
authors.initials: D
authors.lastname: Butler
authors.forename: James H
authors.initials: JH
authors.lastname: Youell
authors.forename: Amanda A
authors.initials: AA
authors.lastname: Brindley
authors.forename: Steve P
authors.initials: SP
authors.lastname: Wood
authors.forename: Martin J
authors.initials: MJ
authors.lastname: Warren
authors.forename: Peter M
authors.initials: PM
authors.lastname: Shoolingin-Jordan
authors.forename: Jonathan B
authors.initials: JB
authors.lastname: Cooper
mesh_terms: D018360:Crystallography, X-Ray; D008958:Models, Molecular; D000623:Porphobilinogen Synthase / Q000737:chemistry* / Q000378:metabolism; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology
chemical_list: D011994:Recombinant Proteins; D000623:Porphobilinogen Synthase