Structure-based rational design of a phosphotriesterase.
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.
Jackson
Colin J
CJ
CSIRO Entomology, Canberra, Australia.
Weir
Kahli
K
Herlt
Anthony
A
Khurana
Jeevan
J
Sutherland
Tara D
TD
Horne
Irene
I
Easton
Christopher
C
Russell
Robyn J
RJ
Scott
Colin
C
Oakeshott
John G
JG
D019943:Amino Acid Substitution / Q000235:genetics*; D020134:Catalytic Domain / Q000235:genetics*; D002709:Chlorfenvinphos / Q000378:metabolism*; D008958:Models, Molecular; D016297:Mutagenesis, Site-Directed; D044345:Phosphoric Triester Hydrolases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism*; D017434:Protein Structure, Tertiary; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012231:Rhizobium / Q000201:enzymology* / Q000235:genetics*; D013379:Substrate Specificity
D011994:Recombinant Proteins; D044345:Phosphoric Triester Hydrolases; D002709:Chlorfenvinphos