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Assay System for N-acylethanolamines Degradation Enzyme, N-acylethanolamine-hydrolyzing Acid Amidase
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  • Assay System for N-acylethanolamines Degradation Enzyme, N-acylethanolamine-hydrolyzing Acid Amidase
  • Assay System for N-acylethanolamines Degradation Enzyme, N-acylethanolamine-hydrolyzing Acid Amidase
저자명
Kim. Dae-Woong,Kim. Gun-Joong,Kim. Hae-Jo,Ghil. Sung-Ho
간행물명
Journal of experimental & biomedical sciences
권/호정보
2012년|18권 4호|pp.438-444 (7 pages)
발행정보
대한의생명과학회
파일정보
정기간행물|ENG|
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이 논문은 한국과학기술정보연구원과 논문 연계를 통해 무료로 제공되는 원문입니다.
서지반출

기타언어초록

N-acylethanolamines (NAEs) including endocannabinoids, anadamide, are long chain fatty acid ethanolamines and express ubiquitously in animal and plant tissues. NAEs have several pharmacological effects including anti-inflammatory, analgesic and anorexic effects. The levels of NAEs in tissues are strictly regulated by synthesizing and hydrolyzing enzymes because NAEs are not stored in the cell but rather made on demand. NAEs are hydrolyzed to free fatty acids and ethanolamines by fatty acid amide hydrolase and N-acylethanolamine-hydrolyzing acid amidase (NAAA). Here, we suggest the fluorescence-based assay system for NAAA. We developed N-(4-methy-2-oxo-2H-chromen-7-yl)palmitamide (PAAC) as a fluorogenic substrate for NAAA and we also generated NAAA stably expressing COSM6 cell line. When extracts of cells expressing NAAA were incubated with PAAC, NAAA specifically hydrolyzed PAAC to palmitic acids and fluorogenic dye, coumarin. Release of coumarin was monitored by using fluorometer. NAAA hydrolyzed PAAC with an apparent Km of $20.05{mu}M$ and Vmax of 32.18 pmol/mg protein/min. This assay system can be used to develop inhibitors or activators of NAAA.