2ps6: Difference between revisions

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[[Image:2ps6.jpg|left|200px]]
{{Seed}}
[[Image:2ps6.png|left|200px]]


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{{STRUCTURE_2ps6|  PDB=2ps6  |  SCENE=  }}  
{{STRUCTURE_2ps6|  PDB=2ps6  |  SCENE=  }}  


'''N225D/S229T trichodiene synthase'''
===N225D/S229T trichodiene synthase===




==Overview==
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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.
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{{ABSTRACT_PUBMED_17996718}}


==About this Structure==
==About this Structure==
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[[Category: Site-directed mutagenesis]]
[[Category: Site-directed mutagenesis]]
[[Category: Terpenoid synthase fold]]
[[Category: Terpenoid synthase fold]]
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Revision as of 23:20, 27 July 2008

File:2ps6.png

Template:STRUCTURE 2ps6

N225D/S229T trichodiene synthase

Template:ABSTRACT PUBMED 17996718

About this Structure

2PS6 is a Single protein structure of sequence from Fusarium sporotrichioides. Full crystallographic information is available from OCA.

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

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