Ueda N, Kurahashi Y, Yamamoto K, Yamamoto S, Tokunaga T
Department of Biochemistry, Tokushima University, School of Medicine, Japan.
J Lipid Mediat Cell Signal. 1996 Sep;14(1-3):57-61. doi: 10.1016/0929-7855(96)00509-3.
Anandamide is an endogenous ligand for cannabinoid receptors. We tried to isolate and purify "anandamide amidohydrolase' which hydrolyzes anandamide to arachidonic acid and ethanolamine. The enzyme activity was found in the microsomal fraction of porcine brain homogenate. The enzyme was solubilized in 1% Triton X-100, and partially purified by hydrophobic chromatography to a specific activity of about 0.3 mumol/min per mg protein (37 degrees C). Apparent K(m) for anandamide was about 60 microM. The enzyme reacted also with ethanolamides of linoleic, oleic, and palmitic acids at lower rates. This enzyme preparation also converted arachidonic acid to anandamide in the presence of 250 mM concentration of ethanolamine. Several lines of evidence including experiments using various inhibitors suggested that the anandamide synthase and amidohydrolase activities were derived from a single enzyme protein.
花生四烯酸乙醇胺是大麻素受体的内源性配体。我们试图分离和纯化“花生四烯酸乙醇胺酰胺水解酶”，该酶可将花生四烯酸乙醇胺水解为花生四烯酸和乙醇胺。在猪脑匀浆的微粒体部分发现了该酶的活性。该酶在1% Triton X-100中溶解，并通过疏水色谱法部分纯化，其比活性约为0.3 μmol/（min·mg蛋白质）（37℃）。花生四烯酸乙醇胺的表观K（m）约为60 μM。该酶也能以较低的速率与亚油酸、油酸和棕榈酸的乙醇酰胺发生反应。在250 mM乙醇胺浓度存在的情况下，这种酶制剂还能将花生四烯酸转化为花生四烯酸乙醇胺。包括使用各种抑制剂的实验在内的几条证据表明，花生四烯酸乙醇胺合酶和酰胺水解酶活性源自单一酶蛋白。
