2ps8: Difference between revisions

Revision as of 19:32, 21 February 2008

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. 2008 Jan 15;469(2):184-94. Epub 2007 Oct 30. PMID:17996718

Page seeded by OCA on Thu Feb 21 18:32:29 2008

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OCA

@@ Line 4: / Line 4: @@
 ==Overview==
-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.
+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 Structure==
@@ Line 10: / Line 10: @@
 ==Reference==
-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:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17996718 17996718]
+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. 2008 Jan 15;469(2):184-94. Epub 2007 Oct 30. PMID:[http://ispc.weizmann.ac.il//pmbin/getpm?pmid=17996718 17996718]
 [[Category: Fusarium sporotrichioides]]
 [[Category: Single protein]]
 [[Category: Trichodiene synthase]]
-[[Category: Cane, D.E.]]
+[[Category: Cane, D E.]]
-[[Category: Christianson, D.W.]]
+[[Category: Christianson, D W.]]
-[[Category: Vedula, L.S.]]
+[[Category: Vedula, L S.]]
 [[Category: EDO]]
 [[Category: MG]]
@@ Line 27: / Line 27: @@
 [[Category: terpenoid synthase fold]]
-''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Wed Jan 23 11:02:09 2008''
+''Page seeded by [http://oca.weizmann.ac.il/oca OCA ] on Thu Feb 21 18:32:29 2008''