Biomedical Science Letters (대한의생명과학회지)

The Korean Society for Biomedical Laboratory Sciences (대한의생명과학회)
권호 표지

Fluorescence-based Assay System for Endocannabinoid Degradation Enzyme, Fatty Acid Amide Hydrolase

  • Kim, Dae-Woong (Department of Life Science, Kyonggi University) ;
  • Kim, Gun-Joong (Department of Chemistry, Hankuk University of Foreign Studies) ;
  • Kim, Hae-Jo (Department of Chemistry, Hankuk University of Foreign Studies) ;
  • Ghil, Sung-Ho (Department of Life Science, Kyonggi University)
  • ;
  • ;
  • ;
  • 길성호 (경기대학교 생명과학과)
  • Received : 2010.10.29
  • Accepted : 2010.12.10
  • Published : 2010.12.31
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Abstract

Endogenous cannabinoids (endocannabinoids) display various pharmacological effects including pain control, anti-inflammation, and neuroprotection. The synthesis and release of endocannabinoids are regulated under both physiological and pathological conditions. The main degrading enzyme of endocannabinoid is fatty acid amide hydrolase (FAAH). Therefore we have developed the fluorescence-based assay system for FAAH. We established stable CosM6 cell lines expressing human FAAH. We also synthesized 2-oxo-2H-chromen-7-yl decanoate (DAEC) as a fluorogenic substrate for FAAH. When crude membrane extracts stably expressing FAAH was incubated with DAEC at $25^{\circ}C$, FAAH reacted specifically to DAEC and catalyzes the hydrolysis of DAEC into decanoic acid and highly fluorescent coumarin. Furthermore, the serin hydrolase inhibitor, phenylmethanesulfonylfluoride, inhibited the coumarin release to the reaction buffer in concentration dependent manner. This assay system is suitable for high-throughput screening since this system has simple experimental procedure and measurement method.

Keywords

References

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