Yang S D, Yu J S, Liu J S, Tzen T C, Wang J K
Biochem Biophys Res Commun. 1987 Jan 15;142(1):38-46. doi: 10.1016/0006-291x(87)90448-7.
Although the activating factor FA of the type-1 protein phosphatase has long been recognized as a cytosolic enzyme involved in the regulation of cell metabolism and nervous functions, strong indications have been obtained that FA is in fact a membrane-bound protein kinase in most mammalian tissues. For instance, direct treatment of the tissue extracts including brain, liver, cardiac, smooth and skeletal muscles with 1% Triton X-100 can cause several fold stimulation of the FA activity. Moreover, at least 50% of the FA can be detected in the particulate fractions of the extracts. Chromatography of the extracts in the presence and absence of Triton X-100 further demonstrate these results. The data can now explain the reason why most people can not isolate reasonable amount of FA from mammalian tissues. It is recommended that Triton X-100 should be used for purification of FA from most mammalian tissue extracts. The results also suggest that most previous studies on the action of FA involved in the regulation of cell functions should be re-evaluated and the membrane-associated FA should be taken into consideration.
尽管1型蛋白磷酸酶的激活因子FA长期以来一直被认为是一种参与细胞代谢和神经功能调节的胞质酶,但已有有力迹象表明,在大多数哺乳动物组织中,FA实际上是一种膜结合蛋白激酶。例如,用1% Triton X-100直接处理包括脑、肝、心脏、平滑肌和骨骼肌在内的组织提取物,可使FA活性提高几倍。此外,在提取物的颗粒部分可检测到至少50%的FA。在有和没有Triton X-100的情况下对提取物进行色谱分析进一步证实了这些结果。这些数据现在可以解释为什么大多数人无法从哺乳动物组织中分离出适量的FA。建议使用Triton X-100从大多数哺乳动物组织提取物中纯化FA。结果还表明,以前大多数关于FA参与细胞功能调节作用的研究都应重新评估,并且应考虑与膜相关的FA。