PURIFICATION, CHAIN SEPARATION AND SEQUENCE OF THE MRC OX-8 ANTIGEN, A MARKER OF RAT CYTO-TOXIC LYMPHOCYTES-T
PURIFICATION, CHAIN SEPARATION AND SEQUENCE OF THE MRC OX-8 ANTIGEN, A MARKER OF RAT CYTO-TOXIC LYMPHOCYTES-T
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DOI：
10.1002/j.1460-2075.1985.tb03968.x
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发表时间：
1985-01-01
期刊：
影响因子：
11.4
通讯作者：
WILLIAMS, AF
中科院分区：
文献类型：
--
作者：
JOHNSON, P;GAGNON, J;WILLIAMS, AF
MRC OX-8抗原是大鼠细胞毒性T淋巴细胞的标志物，由二硫键连接的分子链组成。wts. 37和32 kD。它被认为是等同于人T8和小鼠Lyt 2，3抗原（现在都称为CD 8抗原）。使用单克隆抗体柱从胸腺细胞中纯化MRC OX-8抗原，由于还原和烷基化后保留了抗原性，因此可以通过随后的亲和色谱步骤分离两条多肽链。从每条链中分离肽并测定其序列。将编码小鼠CD 8抗原的cDNA探针（由Dr L.A. Herzenberg）获得大鼠cDNA克隆，从其确定等效大鼠分子的序列。在该翻译序列中鉴定了来自32-kd链的肽，而没有鉴定来自37-kd链的肽。32 kd的多肽序列由210个氨基酸组成，并有一个可能的N-连接糖基化位点。该序列的N-末端部分与小鼠和人类的对应序列惊人地不同，但与其他两个物种一样，它显示出与IG V结构域的明确关系。
The MRC OX-8 antigen is a marker of the rat cytotoxic T lymphocytes that consists of disulphide-linked chains of mol. wts. 37 and 32 kd. It is thought to be equivalent to the human T8 and mouse Lyt2,3 antigens (all now called CD8 antigens). MRC OX-8 antigen was purified from thymocytes using a monoclonal antibody column and because antigenicity was retained after reduction and alkylation the two polypeptide chains could be separated by a subsequent affinity chromatography step. Peptides were isolated from each chain and their sequences determined. A cDNA probe coding for the mouse CD8 antigen (pLY2C-1 provided by Dr L.A. Herzenberg) was used to obtain rat cDNA clones from which the sequence of the equivalent rat molecule was determined. Peptides from the 32-kd chain were identified in this translated sequence whereas peptides from the 37-kd chain were not. The 32-kd polypeptide sequence consisted of 210 amino acids and had one possible N-linked glycosylation site. The N-terminal part of the sequence was surprisingly different from both its mouse and human counterparts but, as in the other two species, it showed a clear relationship to Ig V domains.