title: Trichodiene synthase. Substrate specificity and inhibition.
abstract: The substrate specificity of the sesquiterpene synthase trichodiene synthase was examined by determining the Vmax and Km parameters for the natural substrate, trans,trans-farnesyl diphosphate (1), its stereoisomer, cis,trans-farnesyl diphosphate, and the tertiary allylic isomer, (3R)-nerolidyl diphosphate (3), using both the native fungal and recombinant enzymes. A series of farnesyl diphosphate analogs, 15, 16, 20, 7, 8, and 9, was also tested as inhibitors of trichodiene synthase. 10-Fluorofarnesyl diphosphate (15) was the most effective competitive inhibitor, with a K1 of 16 nM compared to the Km for 1 of 87 nM, while the ether analog of farnesyl diphosphate, 8, an extremely potent inhibitor of squalene synthase, showed only modest inhibition of trichodiene synthase, with a K1/Km of 70.
authors.affiliation: Department of Chemistry, Brown University, Providence, Rhode Island 02912.
authors.forename: D E
authors.initials: DE
authors.lastname: Cane
authors.forename: G
authors.initials: G
authors.lastname: Yang
authors.forename: Q
authors.initials: Q
authors.lastname: Xue
authors.forename: J H
authors.initials: JH
authors.lastname: Shim
mesh_terms: D001667:Binding, Competitive; D019755:Carbon-Carbon Lyases; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005670:Fusarium / Q000201:enzymology / Q000235:genetics; D007700:Kinetics; D008190:Lyases / Q000037:antagonists & inhibitors* / Q000235:genetics / Q000378:metabolism*; D009682:Magnetic Resonance Spectroscopy; D015394:Molecular Structure; D011106:Polyisoprenyl Phosphates / Q000138:chemical synthesis / Q000737:chemistry; D012717:Sesquiterpenes; D013237:Stereoisomerism; D013379:Substrate Specificity
chemical_list: D011106:Polyisoprenyl Phosphates; D012717:Sesquiterpenes; C004808:farnesyl pyrophosphate; D008190:Lyases; D019755:Carbon-Carbon Lyases; C051130:trichodiene synthetase