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Fluorescence-based Assay System for Endocannabinoid Degradation Enzyme, Fatty Acid Amide Hydrolase
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  • Fluorescence-based Assay System for Endocannabinoid Degradation Enzyme, Fatty Acid Amide Hydrolase
  • Fluorescence-based Assay System for Endocannabinoid Degradation Enzyme, Fatty Acid Amide Hydrolase
저자명
길성호,Ghil. Sung-Ho
간행물명
Journal of experimental & biomedical sciences
권/호정보
2010년|16권 4호|pp.279-285 (7 pages)
발행정보
대한의생명과학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
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기타언어초록

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.