Antibodies of predetermined specificity for the NH2 terminus of a cellular protein p53 react with the native molecule: evidence for the presence of different p53s.
Antibodies of predetermined specificity for the NH2 terminus of a cellular protein p53 react with the native molecule: evidence for the presence of different p53s.
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对细胞蛋白 p53 的 NH2 末端具有预定特异性的抗体与天然分子发生反应：不同 p53 存在的证据。
DOI：
10.1073/pnas.80.5.1199
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发表时间：
1983
影响因子：
11.1
通讯作者：
Lerner,R
中科院分区：
文献类型：
--
作者：
Luka,J;Sternås,L;Jörnvall,H;Klein,G;Lerner,R
分别对应于一种称为“p53”的细胞蛋白的残基1-20和10-20的两种合成肽已经与载体蛋白连接，并注射到兔子体内以产生抗体。所获得的抗体能够与天然蛋白质反应，如通过酶联免疫吸附测定、NaDodSO 4凝胶电泳后的免疫印迹的蛋白质A-联染色和免疫沉淀所判断的。这些抗体对蛋白质的免疫测定滴度低于用纯化的p53免疫的抗血清。然而，用免疫印迹方法染色显示，抗p53的抗肽抗体是唯一特异性的。这些数据表明，至少存在两种不同类型的p53分子。细胞蛋白先前分离的人细胞转化的EB病毒和小鼠肿瘤诱导甲基胆蒽似乎是大于p53报告的猿猴病毒40或腺病毒转化的细胞和其他一些肿瘤。一些相互关系尚未被排除，但很明显，这两种蛋白质分子的行为并不相同。抗肽抗体与完整蛋白质的反应对蛋白质构象有影响。此类抗体的严格特异性允许产生可用于定量、纯化和克隆的不同试剂组。
Two synthetic peptides corresponding to residues 1-20 and 10-20, respectively, of one type of a cellular protein called "p53" have been linked to a carrier protein and injected into rabbits to raise antibodies. The antibodies obtained were capable of reacting with the native protein, as judged by an enzyme-linked immunosorbent assay, protein A-linked staining of immunoblots after NaDodSO4 gel electrophoresis, and immunoprecipitation. The immunoassay titers against the protein were lower for these antibodies than for antisera derived from immunization with purified p53. However, staining with the immunoblot method showed that the antipeptide antibodies against p53 were uniquely specific. The data suggest that at least two different types of p53 molecules occur. The cellular protein previously isolated from human cells transformed by Epstein-Barr virus and from murine tumors induced by methylcholanthrene appears to be larger than the p53 reported in relation to simian virus 40- or adenovirus-transformed cells and to some other tumors. Some interrelationships have not been excluded, but it is clear that the two protein molecules do not behave identically. The reactions of the antipeptide antibodies with the intact protein have implications in regard to protein conformations. The strict specificities of such antibodies allow the generation of distinct sets of reagents useful for quantitation, purification, and cloning.