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首页> 外文期刊>Life sciences >ARACHIDONOYL ETHANOLAMIDE-[1,2-C-14] AS A SUBSTRATE FOR ANANDAMIDE AMIDASE
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ARACHIDONOYL ETHANOLAMIDE-[1,2-C-14] AS A SUBSTRATE FOR ANANDAMIDE AMIDASE

机译：花生四烯酸乙醇酰胺-[1,2-C-14]作为花生酰胺酰胺的底物

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Arachidonoyl ethanolamide-[1,2-C-14] was prepared and evaluated as a substrate for anandamide amidase in a radioenzymatic assay that does not require a thin layer chromatography separation step. Using this substrate the release of ethanolamine-[1,2-C-14] is linear for approximately thirty minutes. Anandamide amidase exhibits maximal activity between pH 8 and pH 9 with a steep decline in activity at pH values below 6 and above 10. Arachidonoyl ethanolamide-[1,2- C-14] was used for the assay of anandamide amidase from 10 mu g to 100 mu g protein, from cow brain homogenate, in a 0.2 ml incubation mixture. When plotted as a rectangular hyperbola of the steady-state Michaelis-Menten equation, an approximate K-m of 30 +/- 7 mu M and a V-max of 198 +/- 13 nmoles ethanolamine formed per hour per mg protein homogenate was obtained. [References: 16]
机译：制备了花生四烯酰基乙醇酰胺-[1,2-C-14]，并在不需要薄层色谱分离步骤的放射酶法中，将其作为花生四烯酰胺酰胺酶的底物进行了评估。使用该底物，乙醇胺-[1,2-C-14]的释放线性发生约三十分钟。花生四烯酰胺酰胺酶在pH 8和pH 9之间显示最大活性，在pH值低于6和高于10时活性急剧下降。花生四烯酰乙醇酰胺-[1,2-C-14]用于分析10 µg的花生四烯酰胺酰胺酶在0.2 ml的孵育混合物中，从牛脑匀浆中提取100 µg蛋白。当绘制为稳态Michaelis-Menten方程的矩形双曲线时，每小时每毫克蛋白质匀浆形成的K-m约为30 +/- 7μM，V-max每小时形成198 +/- 13 nmoles乙醇胺。 [参考：16]

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