Purification and properties of an esterase from organophosphate-resistant strain of the mosquito Culex quinquefasciatus.
Organophosphate-resistant and -susceptible strains of Culex quinquefasciatus (mosquito) have been compared on the basis of their esterase activities. The homozygous resistant strain (Dar) shows two highly active esterases after starch-gel electrophoresis, of Rm 0.2 and 0.4, which are absent from susceptible strains (Apo, Mon), and which previous selection studies have shown to be inseparable from organophosphate resistance. After SDS/polyacrylamide-gel electrophoresis and silver staining of total C. quinquefasciatus proteins, a 62 kDa band is observed in strain Dar at high concentrations, and in susceptible strains in trace amounts. After Western blotting, this 62 kDa protein is recognized by antisera raised against the two esterases eluted from starch gels. After chromatofocusing of Dar proteins, the 62 kDa protein is seen to be associated with esterase activity, and of a similar pI to that observed for esterases after isoelectric focusing. Post-translational modification is not required for recognition of the 62 kDa putative esterase, since the protein is immunoprecipitated by the anti-esterase serum from products of translation of Dar mRNA in vitro.
Merryweather
A T
AT
Wolfson Unit of Molecular Genetics, Liverpool School of Tropical Medicine, U.K.
Crampton
J M
JM
Townson
H
H
D000818:Animals; D015153:Blotting, Western; D003465:Culex / Q000187:drug effects / Q000201:enzymology* / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D004591:Electrophoresis, Polyacrylamide Gel; D004592:Electrophoresis, Starch Gel; D004950:Esterases / Q000302:isolation & purification* / Q000378:metabolism; D006720:Homozygote; D006863:Hydrogen-Ion Concentration; D007163:Immunosorbent Techniques; D007306:Insecticides / Q000494:pharmacology*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D009943:Organophosphorus Compounds; D012333:RNA, Messenger / Q000378:metabolism
D007306:Insecticides; D009943:Organophosphorus Compounds; D012333:RNA, Messenger; D004950:Esterases