title: The Schiff base complex of yeast 5-aminolaevulinic acid dehydratase with laevulinic acid.
abstract: The X-ray structure of the complex formed between yeast 5-aminolaevulinic acid dehydratase (ALAD) and the inhibitor laevulinic acid has been determined at 2.15 A resolution. The inhibitor binds by forming a Schiff base link with one of the two invariant lysines at the catalytic center: Lys263. It is known that this lysine forms a Schiff base link with substrate bound at the enzyme's so-called P-site. The carboxyl group of laevulinic acid makes hydrogen bonds with the side-chain-OH groups of Tyr329 and Ser290, as well as with the main-chain >NH group of Ser290. The aliphatic moiety of the inhibitor makes hydrophobic interactions with surrounding aromatic residues in the protein including Phe219, which resides in the flap covering the active site. Our analysis strongly suggests that the same interactions will be made by P-side substrate and also indicates that the substrate that binds at the enzyme's A-site will interact with the enzyme's zinc ion bound by three cysteines (133, 135, and 143). Inhibitor binding caused a substantial ordering of the active site flap (residues 217-235), which was largely invisible in the native electron density map and indicates that this highly conserved yet flexible region has a specific role in substrate binding during catalysis.
authors.affiliation: Division of Biochemistry and Molecular Biology, School of Biological Sciences, University of Southampton, United Kingdom.
authors.forename: P T
authors.initials: PT
authors.lastname: Erskine
authors.forename: R
authors.initials: R
authors.lastname: Newbold
authors.forename: J
authors.initials: J
authors.lastname: Roper
authors.forename: A
authors.initials: A
authors.lastname: Coker
authors.forename: M J
authors.initials: MJ
authors.lastname: Warren
authors.forename: P M
authors.initials: PM
authors.lastname: Shoolingin-Jordan
authors.forename: S P
authors.initials: SP
authors.lastname: Wood
authors.forename: J B
authors.initials: JB
authors.lastname: Cooper
mesh_terms: D001665:Binding Sites; D002384:Catalysis; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D007982:Levulinic Acids / Q000737:chemistry*; D000623:Porphobilinogen Synthase / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000201:enzymology*; D012545:Schiff Bases / Q000737:chemistry*
chemical_list: D007982:Levulinic Acids; D012545:Schiff Bases; D000623:Porphobilinogen Synthase; C032246:levulinic acid