Development of fluorescent substrates for microsomal epoxide hydrolase and application to inhibition studies

Christophe Morisseau, Maud Bernay, Aurélie Escaich, James R. Sanborn, Jozsef Lango, Bruce D. Hammock

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

The microsomal epoxide hydrolase (mEH) plays a significant role in the metabolism of numerous xenobiotics. In addition, it has a potential role in sexual development and bile acid transport, and it is associated with a number of diseases such as emphysema, spontaneous abortion, eclampsia, and several forms of cancer. Toward developing chemical tools to study the biological role of mEH, we designed and synthesized a series of absorbent and fluorescent substrates. The highest activity for both rat and human mEH was obtained with the fluorescent substrate cyano(6-methoxy-naphthalen-2-yl)methyl glycidyl carbonate (11). An in vitro inhibition assay using this substrate ranked a series of known inhibitors similarly to the assay that used radioactive cis-stilbene oxide but with a greater discrimination between inhibitors. These results demonstrate that the new fluorescence-based assay is a useful tool for the discovery of structure-activity relationships among mEH inhibitors. Furthermore, this substrate could also be used for the screening chemical library with high accuracy and with a Z′ value of approximately 0.7. This new assay permits a significant decrease in labor and cost and also offers the advantage of a continuous readout. However, it should not be used with crude enzyme preparations due to interfering reactions.

Original languageEnglish (US)
Pages (from-to)154-162
Number of pages9
JournalAnalytical Biochemistry
Volume414
Issue number1
DOIs
StatePublished - Jul 1 2011

Fingerprint

Epoxide Hydrolases
Assays
Substrates
Small Molecule Libraries
Eclampsia
Sexual Development
Carbonates
Emphysema
Spontaneous Abortion
Xenobiotics
Structure-Activity Relationship
Bile Acids and Salts
Metabolism
Rats
Screening
Fluorescence
Personnel
Costs and Cost Analysis
Enzymes
Costs

Keywords

  • α-Cyanocarbonate
  • Fluorescent substrate
  • Microsomal epoxide hydrolase
  • Xenobiotic metabolism

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

Development of fluorescent substrates for microsomal epoxide hydrolase and application to inhibition studies. / Morisseau, Christophe; Bernay, Maud; Escaich, Aurélie; Sanborn, James R.; Lango, Jozsef; Hammock, Bruce D.

In: Analytical Biochemistry, Vol. 414, No. 1, 01.07.2011, p. 154-162.

Research output: Contribution to journalArticle

Morisseau, C, Bernay, M, Escaich, A, Sanborn, JR, Lango, J & Hammock, BD 2011, 'Development of fluorescent substrates for microsomal epoxide hydrolase and application to inhibition studies', Analytical Biochemistry, vol. 414, no. 1, pp. 154-162. https://doi.org/10.1016/j.ab.2011.02.038
Morisseau, Christophe ; Bernay, Maud ; Escaich, Aurélie ; Sanborn, James R. ; Lango, Jozsef ; Hammock, Bruce D. / Development of fluorescent substrates for microsomal epoxide hydrolase and application to inhibition studies. In: Analytical Biochemistry. 2011 ; Vol. 414, No. 1. pp. 154-162.
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