Grussenmeyer T, Scheidtmann K H, Hutchinson M A, Eckhart W, Walter G
Proc Natl Acad Sci U S A. 1985 Dec;82(23):7952-4. doi: 10.1073/pnas.82.23.7952.
Antibodies against synthetic peptides corresponding to the carboxyl-terminal six amino acids, Lys-Arg-Ser-Arg-His-Phe (KF), and an internal region, Glu-Glu-Glu-Glu-Tyr-Met-Pro-Met-Glu (EE), of polyoma virus medium T antigen were used successively to purify medium T antigen by affinity chromatography. Medium T antigen from cell extracts was first bound to anti-KF antibodies and released from the immune complex with excess KF peptide; then it was bound to anti-EE antibodies and released with excess EE peptide. Two proteins, pp60c-src and a new protein of approximately equal to 61,000 Da (61-kDa protein), were copurified because they formed complexes with medium T antigen. The 61-kDa protein-medium T antigen complex was detected in extracts from wild-type-infected and transformed cells but not from cells infected with NG59 virus, which has a mutation in the medium T gene and is transformation defective. Instead, NG59 medium T antigen formed a complex with another cellular protein of approximately equal to 72,000 Da.
利用针对多瘤病毒中T抗原羧基末端六个氨基酸（Lys-Arg-Ser-Arg-His-Phe，KF）以及内部区域（Glu-Glu-Glu-Glu-Tyr-Met-Pro-Met-Glu，EE）的合成肽的抗体，通过亲和层析依次纯化中T抗原。细胞提取物中的中T抗原首先与抗KF抗体结合，然后用过量的KF肽从免疫复合物中释放出来；接着它与抗EE抗体结合，再用过量的EE肽释放出来。两种蛋白质，pp60c-src和一种约61,000 Da的新蛋白质（61-kDa蛋白质）被共纯化，因为它们与中T抗原形成了复合物。在野生型感染和转化细胞的提取物中检测到了61-kDa蛋白质-中T抗原复合物，但在感染了NG59病毒的细胞中未检测到，NG59病毒的中T基因发生了突变且转化缺陷。相反，NG59中T抗原与另一种约72,000 Da的细胞蛋白质形成了复合物。
