Premium
An acid amidase hydrolyzing anandamide as an endogenous ligand forcannabinoid receptors
Author(s) -
Natsuo Ueda,
Kazushi Yamanaka,
Yuka Terasawa,
Shozo Yamamoto
Publication year - 1999
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/s0014-5793(99)00820-0
Subject(s) - anandamide , amidase , cannabinoid receptor , ligand (biochemistry) , endocannabinoid system , receptor , citation , chemistry , information retrieval , computational biology , biochemistry , computer science , biology , world wide web , enzyme , agonist
Anandamide loses its cannabimimetic activities upon hydrolysis to arachidonic acid and ethanolamine. So far the anandamide hydrolyzing activity widely distributed in mammalian organs has been attributed exclusively to an enzyme referred to as anandamide amidohydrolase with an optimum pH around 9. We found another enzyme hydrolyzing anandamide in a human megakaryoblastic cell line (CMK). The enzyme present in the 12,000 x g pellet of the cell homogenate was solubilized by freeze-thaw. The solubilized enzyme showed an optimal pH around 5, and was almost inactive at alkaline pH. The enzyme activity was increased by the addition of dithiothreitol. In contrast, anandamide amidohydrolase of RBL-1 cells was mostly insoluble even after freeze-thaw, showed an optimal pH at 9, and was not affected by dithiothreitol. Furthermore, the enzyme of CMK cells was much less sensitive to phenylmethylsulfonyl fluoride and methyl arachidonoyl fluorophosphonate potently inhibiting anandamide amidohydrolase, and effectively hydrolyzed palmitoylethanolamide, which was a poor substrate for anandamide amidohydrolase. Thus, the enzyme of CMK cells is distinguishable from anandamide amidohydrolase.