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本文介绍腺苷酸环化酶(AC)和环腺替酸磷酸二酯酶(PDE)比活力的一种简单、快速测定法。以腺苷三磷酸(ATP)或环磷酸腺苷(cAMP)作为底物,同肝匀浆孵育时生成的或未消耗的 cAMP,用氧化铝柱层析法分离,以 Tris-HCl 缓冲液(pH7.4)洗脱,用分光光度法测定洗脱液中的 cAMP。根据 cAMP 的生成量和 cAMP 的消耗量分别表示 AC 和 PDE 活力。测定了小白鼠肝组织中 AC 和 PDE 比活力。上清液和沉淀中 AC 比活力分别为0.96和1.64单位/mg 蛋白质、PDE 比活力分别为2.06和0.50单位/mg 蛋白质,上清中 AC/PDE 比值为0.47,沉淀中比值为3.30。
This article describes a simple, rapid assay for the specific activities of adenylate cyclase (AC) and cyclohumginate phosphodiesterase (PDE). The cAMP produced or not consumed when incubated with liver homogenate with adenosine triphosphate (ATP) or cAMP as a substrate was separated by alumina column chromatography and eluted with Tris-HCl buffer ( pH 7.4), and the cAMP in the eluate was measured spectrophotometrically. The AC and PDE activities are expressed respectively based on the amount of cAMP produced and the consumption of cAMP. The specific activities of AC and PDE in the liver of mice were measured. The specific activity of AC in supernatant and pellet were 0.96 and 1.64 units / mg protein respectively, and the specific activity of PDE was 2.06 and 0.50 units / mg protein respectively. The ratio of AC / PDE in the supernatant was 0.47 and the ratio in sediment was 3.30.