1. Structure-based rational design of a phosphotriesterase
- Author
-
Irene Horne, Jeevan L. Khurana, Colin Scott, Colin J. Jackson, Christopher J. Easton, Khali Weir, Anthony J. Herlt, Tara D. Sutherland, John G. Oakeshott, and Robyn J. Russell
- Subjects
Models, Molecular ,Stereochemistry ,In silico ,Applied Microbiology and Biotechnology ,Substrate Specificity ,Hydrolysis ,chemistry.chemical_compound ,Stereospecificity ,Catalytic Domain ,Enzymology and Protein Engineering ,Ecology ,Chemistry ,Rational design ,Chlorfenvinphos ,Substrate (chemistry) ,Recombinant Proteins ,Protein Structure, Tertiary ,Phosphoric Triester Hydrolases ,Amino Acid Substitution ,Mutagenesis, Site-Directed ,Structure based ,Food Science ,Biotechnology ,Agrobacterium radiobacter ,Rhizobium - Abstract
In silico substrate docking of both stereoisomers of the pesticide chlorfenvinphos (CVP) in the phosphotriesterase from Agrobacterium radiobacter identified two residues (F131 and W132) that prevent productive substrate binding and cause stereospecificity. A variant (W131H/F132A) was designed that exhibited ca. 480-fold and 8-fold increases in the rate of Z -CVP and E -CVP hydrolysis, respectively, eliminating stereospecificity.
- Published
- 2009