通过饱和分析测定脑组织中3':5'-环磷酸腺苷。对一种使用牛肌肉结合蛋白的方法的评估。
Determination of adenosine 3':5'-cyclic monophosphate in cerebral tissues by saturation analysis. Assessment of a method using a binding protein from ox muscle.
作者信息
Weller M, Rodnight R, Carrera D
出版信息
Biochem J. 1972 Aug;129(1):113-21. doi: 10.1042/bj1290113.
Abstract
- The Gilman (1970) procedure for determining cyclic AMP (adenosine 3':5'-cyclic monophosphate) by saturation analysis gave erroneous results when applied to the analysis of extracts of whole brain or preparations of membrane fragments from brain. 2. The extracts contained a non-diffusible factor, which enhanced the binding of cyclic AMP by the muscle protein fraction. 3. Extracts also contained material which inhibited binding, but net inhibition of binding was only observed when relatively concentrated extracts were analysed. 4. The error introduced by the factors modifying binding could be eliminated by incorporation of unlabelled internal standards in the unknowns. The design adopted enables a statistical estimate to be made of the standard error of a single assay. 5. The modified assay was used to determine bound cyclic AMP and adenylate cyclase activity in cerebral membrane fragments. Five preparations of synaptic membrane fragments contained less than 3.5pmol of cyclic AMP/mg of protein; a microsomal fraction from rat contained 65pmol of cyclic AMP/mg of protein.
摘要
- 吉尔曼（1970年）采用饱和分析法测定环磷酸腺苷（腺苷3':5'-环磷酸单酯）时，将其应用于全脑提取物或脑细胞膜碎片制剂的分析时得出了错误结果。2. 提取物中含有一种不可扩散因子，它增强了肌肉蛋白组分对环磷酸腺苷的结合。3. 提取物中还含有抑制结合的物质，但只有在分析相对浓缩的提取物时才观察到结合的净抑制。4. 通过在未知样品中加入未标记的内标，可以消除因修饰结合的因子所引入的误差。所采用的设计能够对单次测定的标准误差进行统计估计。5. 改良后的测定法用于测定脑膜碎片中结合的环磷酸腺苷和腺苷酸环化酶活性。五种突触膜碎片制剂每毫克蛋白质所含的环磷酸腺苷少于3.5皮摩尔；大鼠的微粒体组分每毫克蛋白质含有65皮摩尔环磷酸腺苷。
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