Development of pyrethroid-like fluorescent substrates for glutathione S-transferase

Huazhang Huang, Hongwei Yao, Jun Yan Liu, Aman I. Samra, Shizuo G. Kamita, Anthony J. Cornel, Bruce D. Hammock

Research output: Contribution to journalArticle

8 Scopus citations

Abstract

The availability of highly sensitive substrates is critical for the development of precise and rapid assays for detecting changes in glutathione S-transferase (GST) activity that are associated with GST-mediated metabolism of insecticides. In this study, six pyrethroid-like compounds were synthesized and characterized as substrates for insect and mammalian GSTs. All of the substrates were esters composed of the same alcohol moiety, 7-hydroxy-4- methylcoumarin, and acid moieties that structurally mimic some commonly used pyrethroid insecticides, including cypermethrin and cyhalothrin. CpGSTD1, a recombinant Delta class GST from the mosquito Culex pipiens pipiens, metabolized our pyrethroid-like substrates with both chemical and geometric preference (i.e., the cis-isomers were metabolized at 2- to 5-fold higher rates than the corresponding trans-isomers). A GST preparation from mouse liver also metabolized most of our pyrethroid-like substrates with both chemical and geometric preference but at 10- to 170-fold lower rates. CpGSTD1 and mouse GSTs metabolized 1-chloro-2,4-dinitrobenezene (CDNB), a general GST substrate, at more than 200-fold higher rates than our novel pyrethroid-like substrates. There was a 10-fold difference in the specificity constant (kcat/K M ratio) of CpGSTD1 for CDNB and those of CpGSTD1 for cis-DCVC and cis-TFMCVC, suggesting that cis-DCVC and cis-TFMCVC may be useful for the detection of GST-based metabolism of pyrethroids in mosquitoes.

Original languageEnglish (US)
Pages (from-to)77-83
Number of pages7
JournalAnalytical Biochemistry
Volume431
Issue number2
DOIs
StatePublished - Dec 15 2012

Keywords

  • Fluorescent substrate
  • Glutathione S-transferase (GST)
  • Pyrethroid

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

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    Huang, H., Yao, H., Liu, J. Y., Samra, A. I., Kamita, S. G., Cornel, A. J., & Hammock, B. D. (2012). Development of pyrethroid-like fluorescent substrates for glutathione S-transferase. Analytical Biochemistry, 431(2), 77-83. https://doi.org/10.1016/j.ab.2012.09.011