Purchio A F
J Virol. 1982 Jan;41(1):1-7. doi: 10.1128/JVI.41.1.1-7.1982.
pp60src, the product of the rous sarcoma virus src gene, was purified greater than 100,000-fold by a combination of ion-exchange and immunoaffinity chromatography. Incubation of pp60src purified in this fashion with [32P-gamma]ATP resulted in a single 32P-labeled protein when analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Staining of these gels with silver nitrate showed a predominant 60,000-dalton polypeptide which comigrated with the protein labeled with 32P in vitro. Partial digestion of this protein with V8 protease after in vitro iodination indicated that it was pp60src. These results suggest that pp60src is able to autophosphorylate.
pp60src是劳氏肉瘤病毒src基因的产物,通过离子交换和免疫亲和层析相结合的方法纯化了超过10万倍。用[32P-γ]ATP孵育以这种方式纯化的pp60src,经十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分析时产生单一的32P标记蛋白。用硝酸银对这些凝胶进行染色显示,有一条主要的60000道尔顿的多肽,它在体外与用32P标记的蛋白迁移率相同。体外碘化后用V8蛋白酶对该蛋白进行部分消化表明它就是pp60src。这些结果表明pp60src能够自身磷酸化。