Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates
Measurement of Second Messengers in Signal Transduction: cAMP and Inositol Phosphates
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DOI：
10.1002/0471142301.ns0712s05
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发表时间：
1998-11
影响因子：
--
通讯作者：
F. Gusovsky
中科院分区：
文献类型：
--
作者：
F. Gusovsky
cAMP作为激素作用的细胞内介质，并且细胞和组织中cAMP水平的准确定量测定的重要性被广泛认识。本文描述了用于测定膜中腺苷酸环化酶活性的最常用方法，用于测量两步色谱分离后[α-32 P]ATP转化为[32 P]cAMP。在信号转导中也是关键的是磷酸肌醇周转，其与细胞溶质钙浓度变化引起的受体活化有关。如本单元所述，通过用[3 H]-肌醇标记磷脂池，然后分析氚化肌醇磷酸盐，可以测量磷酸肌醇周转率。
cAMP acts as an intracellular mediator of hormone action and the importance of accurate quantitative determination of cAMP levels in cells and tissues is widely recognized. The most utilized procedures for the determination of adenylate cyclase activity in membranes are described here for measuring the conversion of [alpha‐32P]ATP into [32P]cAMP after a two‐step chromatographic separation. Also critical in signal tranduction is phosphoinositide turnover, which is linked to receptor activation resulting from changes in cytosolic calcium concentrations. Phosphoinositide turnover can be measured as described in this unit by labeling phospholipid pools with [3H]‐inositol and then analyzing for tritiated inositol phosphates.