COMPLEXES OF POLYOMA-VIRUS MEDIUM T-ANTIGEN AND CELLULAR PROTEINS
COMPLEXES OF POLYOMA-VIRUS MEDIUM T-ANTIGEN AND CELLULAR PROTEINS
复制标题
DOI：
10.1073/pnas.82.23.7952
复制
发表时间：
1985-01-01
影响因子：
11.1
通讯作者：
WALTER, G
中科院分区：
文献类型：
--
作者：
GRUSSENMEYER, T;SCHEIDTMANN, KH;WALTER, G
用抗多瘤病毒T抗原羧基末端6个氨基酸Lys-Arg-Ser-Arg-His-Phe(KF)和内部区域Glu-Tyr-Met-Pro-Met-Glu(EE)的抗体，亲和层析纯化T抗原。细胞提取液中的T抗原首先与抗KF抗体结合，并与过量的KF多肽一起从免疫复合体中释放出来，然后与抗EE抗体结合，再与过量的EE多肽释放。Pp60c-src和apprxeq的一个新蛋白。61,000 Da(61 kDa蛋白)，由于它们与中等T抗原形成复合物而被融合。在野生型感染和转化细胞的提取液中检测到61 kDa的蛋白-培养基T抗原复合体，而在感染NG59病毒的细胞中未检测到该复合体，NG59病毒的培养基T基因存在突变，是转化缺陷。相反，NG59中T抗原与另一种细胞蛋白.apprxeq形成了一个复合体。72000 Da.
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 .apprxeq. 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 .apprxeq. 72,000 Da.