2ps8
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Y295F Trichodiene Synthase: Complex With Mg and Pyrophosphate
OverviewOverview
Trichodiene synthase from Fusarium sporotrichioides contains two metal, ion-binding motifs required for the cyclization of farnesyl diphosphate:, the "aspartate-rich" motif D(100)DXX(D/E) that coordinates to Mg2+A and, Mg2+C, and the "NSE/DTE" motif N(225)DXXSXXXE that chelates Mg2+B, (boldface indicates metal ion ligands). Here, we report steady-state, kinetic parameters, product array analyses, and X-ray crystal structures, of trichodiene synthase mutants in which the fungal NSE motif is, progressively converted into a plant-like DDXXTXXXE motif, resulting in a, degradation in both steady-state kinetic parameters and product, specificity. Each catalytically active mutant generates a different, distribution of sesquiterpene products, and three newly detected, sesquiterpenes are identified. In addition, the kinetic and structural, properties of the Y295F mutant of trichodiene synthase were found to be, similar to those of the wild-type enzyme, thereby ruling out a proposed, role for Y295 in catalysis.
About this StructureAbout this Structure
2PS8 is a Single protein structure of sequence from Fusarium sporotrichioides with , and as ligands. Active as Trichodiene synthase, with EC number 4.2.3.6 Full crystallographic information is available from OCA.
ReferenceReference
Structural and mechanistic analysis of trichodiene synthase using site-directed mutagenesis: Probing the catalytic function of tyrosine-295 and the asparagine-225/serine-229/glutamate-233-Mg2+B motif., Vedula LS, Jiang J, Zakharian T, Cane DE, Christianson DW, Arch Biochem Biophys. 2007 Oct 30;. PMID:17996718
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