Identification, expression, and purification of a pyrethroidhydrolyzing carboxylesterase from mouse liver microsomes

Stok, Jeanette E., Huang, Huazhang, Jones, Paul D., Wheelock, Craig E., Morisseau, Christopher and Hammock, Bruce D. (2004) Identification, expression, and purification of a pyrethroidhydrolyzing carboxylesterase from mouse liver microsomes. Journal of Biological Chemistry, 279 28: 29863-29869. doi:10.1074/jbc.M403673200

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Author Stok, Jeanette E.
Huang, Huazhang
Jones, Paul D.
Wheelock, Craig E.
Morisseau, Christopher
Hammock, Bruce D.
Title Identification, expression, and purification of a pyrethroidhydrolyzing carboxylesterase from mouse liver microsomes
Journal name Journal of Biological Chemistry   Check publisher's open access policy
ISSN 0021-9258
Publication date 2004-07-09
Year available 2004
Sub-type Article (original research)
DOI 10.1074/jbc.M403673200
Open Access Status File (Publisher version)
Volume 279
Issue 28
Start page 29863
End page 29869
Total pages 7
Place of publication Bethesda, U.S.
Publisher American Society for Biochemistry and Molecular Biology,
Language eng
Subject 03 Chemical Sciences
0304 Medicinal and Biomolecular Chemistry
Abstract Carboxylesterases are enzymes that catalyze the hydrolysis of a wide range of ester-containing endogenous and xenobiotic compounds. Although the use of pyrethroids is increasing, the specific enzymes involved in the hydrolysis of these insecticides have yet to be identified. A pyrethroid-hydrolyzing enzyme was partially purified from mouse liver microsomes using a fluorescent reporter similar in structure to cypermethrin (Shan, G., and Hammock, B. D. (2001) Anal. Biochem. 299, 54-62 and Wheelock, C. E., Wheelock, A. M., Zhang, R., Stok, J. E., Morisseau, C., Le Valley, S. E., Green, C. E., and Hammock, B. D. (2003) Anal. Biochem. 315, 208-222) and subsequently identified as a carboxylesterase (NCBI accession number BAC36707). The expressed sequence tag was then cloned, expressed in baculovirus, and purified to homogeneity. Kinetic constants for a large number of both type I and type II pyrethroid or pyrethroid-like substrates were determined. This esterase possesses similar kinetic constants for cypermethrin and its fluorescent-surrogate (kcat = 0.12 ± 0.03 versus 0.11 ± 0.01 s-1). Compared with their cis- counterparts, trans-permethrin and cypermethrin were hydrolyzed 22- and 4-fold faster, respectively. Of the four fenvalerate isomers the (2R)(αR)-isomer was hydrolyzed at least 1 order of magnitude faster than any other isomer. However, it is unlikely that this enzyme accounts for the total pyrethroid hydrolysis in the microsomes because both isoelectrofocusing and native PAGE indicate the presence of a second region of cypermethrin-metabolizing enzymes. A second carboxylesterase gene (NCBI accession number NM_133960), isolated during a cDNA mouse liver library screening, was also found to hydrolyze pyrethroids. Both these enzymes could be used as preliminary tools in establishing the relative toxicity of new pyrethroids.
Keyword Carboxylesterase
Pyrethroid hydrolysis
Q-Index Code C1

Document type: Journal Article
Sub-type: Article (original research)
Collections: Excellence in Research Australia (ERA) - Collection
School of Chemistry and Molecular Biosciences
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Citation counts: TR Web of Science Citation Count  Cited 37 times in Thomson Reuters Web of Science Article | Citations
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Created: Mon, 21 Dec 2009, 10:43:33 EST by Macushla Boyle on behalf of Faculty of Science