Schraven B, Kirchgessner H, Gaber B, Samstag Y, Meuer S
Abteilung Angewandte Immunologie, Deutsches Krebsforschungszentrum, Heidelberg, FRG.
Eur J Immunol. 1991 Oct;21(10):2469-77. doi: 10.1002/eji.1830211025.
In vitro protein kinase assays of CD45 immunoprecipitates prepared from digitonin lysates of resting human T lymphocytes resulted in exclusive tyrosine phosphorylation of a 32-kDa protein (pp32). Reprecipitation of the in vitro phosphorylated proteins and Western blot analysis of whole CD45 immunoprecipitates employing antisera specifically directed at different protein tyrosine kinases demonstrated that the p56lck protein tyrosine kinase was responsible for in vitro phosphorylation of pp32. Since in vitro kinase assays of p56lck immunoprecipitates also resulted in phosphorylation of pp32, the present data strongly suggest that a functional complex is formed between CD45, p56lck and pp32. Such a notion is supported by the findings that phosphorylation of pp32 by p56lck correlated with expression of the CD45 molecules and that in vitro phosphorylated pp32 was completely dephosphorylated by purified CD45.
对从静息人T淋巴细胞的洋地黄皂苷裂解物制备的CD45免疫沉淀物进行的体外蛋白激酶分析,结果显示一种32 kDa蛋白(pp32)发生了特异性酪氨酸磷酸化。对体外磷酸化蛋白进行再沉淀,并使用针对不同蛋白酪氨酸激酶的抗血清对整个CD45免疫沉淀物进行蛋白质印迹分析,结果表明p56lck蛋白酪氨酸激酶负责pp32的体外磷酸化。由于对p56lck免疫沉淀物进行的体外激酶分析也导致pp32发生磷酸化,目前的数据强烈表明在CD45、p56lck和pp32之间形成了功能复合物。p56lck对pp32的磷酸化与CD45分子的表达相关,以及体外磷酸化的pp32被纯化的CD45完全去磷酸化,这些发现支持了这一观点。