title: The x-ray structure of yeast 5-aminolaevulinic acid dehydratase complexed with substrate and three inhibitors.
abstract: The structures of 5-aminolaevulinic acid dehydratase (ALAD) complexed with substrate (5-aminolaevulinic acid) and three inhibitors: laevulinic acid, succinylacetone and 4-keto-5-aminolaevulinic acid, have been solved at high resolution. The ligands all bind by forming a covalent link with Lys263 at the active site. The structures define the interactions made by one of the two substrate moieties that bind to the enzyme during catalysis. All of the inhibitors induce a significant ordering of the flap covering the active site. Succinylacetone appears to be unique by inducing a number of conformational changes in loops covering the active site, which may be important for understanding the co-operative properties of ALAD enzymes. Succinylacetone is produced in large amounts by patients suffering from the hereditary disease type I tyrosinaemia and its potent inhibition of ALAD also has implications for the pathology of this disease. The most intriguing result is that obtained with 4-keto-5-amino-hexanoic acid, which seems to form a stable carbinolamine intermediate with Lys263. It appears that we have defined the structure of an intermediate of Schiff base formation that the substrate forms upon binding to the P-site of the enzyme.
authors.affiliation: Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, Bassett Crescent East, Southampton SO16 7PX, UK.
authors.forename: P T
authors.initials: PT
authors.lastname: Erskine
authors.forename: R
authors.initials: R
authors.lastname: Newbold
authors.forename: A A
authors.initials: AA
authors.lastname: Brindley
authors.forename: S P
authors.initials: SP
authors.lastname: Wood
authors.forename: P M
authors.initials: PM
authors.lastname: Shoolingin-Jordan
authors.forename: M J
authors.initials: MJ
authors.lastname: Warren
authors.forename: J B
authors.initials: JB
authors.lastname: Cooper
mesh_terms: D000622:Aminolevulinic Acid / Q000737:chemistry / Q000378:metabolism; D001667:Binding, Competitive; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004791:Enzyme Inhibitors / Q000737:chemistry* / Q000378:metabolism; D006537:Heptanoates / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D007982:Levulinic Acids / Q000737:chemistry / Q000378:metabolism; D008239:Lysine / Q000737:chemistry; D008958:Models, Molecular; D000623:Porphobilinogen Synthase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D020176:Tyrosinemias / Q000378:metabolism; D015003:Yeasts / Q000201:enzymology*
chemical_list: C436711:5-amino-4-ketolevulinic acid; D004791:Enzyme Inhibitors; D006537:Heptanoates; D007982:Levulinic Acids; C020804:succinylacetone; D000622:Aminolevulinic Acid; D000623:Porphobilinogen Synthase; D008239:Lysine; C032246:levulinic acid