一种新型磷脂酰肌醇特异性磷脂酶C（β亚家族的第三个成员）的鉴定、纯化及特性分析。
Identification, purification and characterization of a novel phosphatidylinositol-specific phospholipase C, a third member of the beta subfamily.
作者信息
Carozzi A J, Kriz R W, Webster C, Parker P J
机构信息
Imperial Cancer Research Fund, Lincoln's Inn Fields, London, England.
出版信息
Eur J Biochem. 1992 Dec 1;210(2):521-9. doi: 10.1111/j.1432-1033.1992.tb17450.x.
The partial sequence of a novel PtdIns-specific phospholipase C of the beta subfamily (PtdIns-PLC beta 3) is described. Based upon the predicted protein sequence, monospecific antibodies have been raised and used to identify a suitable source for purification of the protein. Fractionation of HeLa S3 cells revealed that immunoreactive PtdIns-PLC beta 3 is membrane associated; purification (approximately 1000-fold) from this fraction yielded a single immunoreactive protein of 158 kDa, with a specific activity of 136 mumol.min-1.mg-1, with PtdIns 4,5-bisphosphate as substrate. Substrate specificity and Ca2+ dependence of this purified PtdIns-PLC are characteristic of the PtdIns-PLC beta subfamily.
描述了一种新型β亚家族磷脂酰肌醇特异性磷脂酶C（PtdIns-PLCβ3）的部分序列。根据预测的蛋白质序列，制备了单特异性抗体，并用于确定纯化该蛋白质的合适来源。对HeLa S3细胞进行分级分离发现，免疫反应性PtdIns-PLCβ3与膜相关；从该级分中纯化（约1000倍）得到一种158 kDa的单一免疫反应性蛋白质，以磷脂酰肌醇4,5-二磷酸为底物时，比活性为136 μmol·min-1·mg-1。这种纯化的PtdIns-PLC的底物特异性和Ca2+依赖性是PtdIns-PLCβ亚家族的特征。
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