Mazurenko N N, Tishchenko V A, Vinogradov V A, Kiselev F L
Mol Biol (Mosk). 1988 May-Jun;22(3):844-52.
To generate the antibodies to the transforming protein of Rous sarcoma virus (RSV) pp60src, rabbits were immunized with the peptide, corresponding to 415-421 sequence of pp60src. These antibodies immunoprecipitate pp60src in RSV-transformed chicken and mammalian cells, and also some proteins (45, 85 and 120 kDa), which could be autophosphorylated in vitro. It was shown that 415-421 sequence of pp60src is not recognized by the antibodies to pp60src from RSV-induced tumour bearing rabbits (TBR serum). In contrast to TBR serum, antibodies, generated against synthetic peptide, corresponding 415-421 sequence of pp60src couldn't be phosphorylated in vitro, when [gamma-32P]ATP is added to the immune complex. The antipeptide antibodies, bound to pp60src did not block phosphorylation of TBR immunoglobulins, added to this immune complex. Hence, 415-421 sequence of pp60src RSV containing the major tyrosine phosphorylation site does not take part in the kinase reaction in vitro.
为了产生针对劳斯肉瘤病毒（RSV）的转化蛋白pp60src的抗体，用对应于pp60src 415 - 421序列的肽对兔子进行免疫。这些抗体能在RSV转化的鸡和哺乳动物细胞中免疫沉淀pp60src，还能沉淀一些在体外可自磷酸化的蛋白质（45、85和120 kDa）。结果表明，RSV诱导的荷瘤兔（TBR血清）产生的针对pp60src的抗体不能识别pp60src的415 - 421序列。与TBR血清不同，针对对应于pp60src 415 - 421序列的合成肽产生的抗体，在向免疫复合物中加入[γ - 32P]ATP时，在体外不能被磷酸化。与pp60src结合的抗肽抗体不会阻断加入该免疫复合物中的TBR免疫球蛋白的磷酸化。因此，含有主要酪氨酸磷酸化位点的pp60src RSV的415 - 421序列不参与体外激酶反应。
