Takahashi M, Buma Y, Taniguchi M
Second Department of Pathology, Nagoya University, School of Medicine, Japan.
Oncogene. 1991 Feb;6(2):297-301.
Monoclonal and/or polyclonal antibodies were generated against the products synthesized from two portions of the ret proto-oncogene (c-ret) cDNA expressed in Escherichia coli. These antibodies were reactive in immunoblotting with 150 kd and 170 kd proteins in cell lysates from three human neuroblastoma cell lines expressing the ret proto-oncogene. When the neuroblastoma cells were treated with tunicamycin, a protein with an apparent molecular weight of 120 kd, which is consistent with that of the c-ret protein predicted from the cDNA sequence, appeared on immunoblots. These results indicated that the 150 kd and 170 kd proteins in neuroblastoma cells are produced from a single polypeptide of 120 kd by posttranslational glycosylation. Furthermore, the antibodies detected a unique 190 kd protein as well as 150 kd protein in a cell lysate from THP-1 human monocytic leukemia cell line, suggesting that glycosylated forms of the c-ret protein are different between neuroblastoma and leukemia cells.
针对在大肠杆菌中表达的原癌基因ret(c-ret)cDNA的两个部分所合成的产物,制备了单克隆抗体和/或多克隆抗体。这些抗体在免疫印迹中,能与来自三种表达ret原癌基因的人神经母细胞瘤细胞系的细胞裂解物中的150kd和170kd蛋白质发生反应。当用衣霉素处理神经母细胞瘤细胞时,免疫印迹上出现了一种表观分子量为120kd的蛋白质,这与从cDNA序列预测的c-ret蛋白质分子量一致。这些结果表明,神经母细胞瘤细胞中的150kd和170kd蛋白质是由120kd的单一多肽经翻译后糖基化产生的。此外,这些抗体在THP-1人单核细胞白血病细胞系的细胞裂解物中检测到一种独特的190kd蛋白质以及150kd蛋白质,这表明神经母细胞瘤细胞和白血病细胞中c-ret蛋白质的糖基化形式有所不同。