A monoclonal antibody reactive with an activated ras protein expressing valine at position 12
A monoclonal antibody reactive with an activated ras protein expressing valine at position 12
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与 12 位表达缬氨酸的激活 ras 蛋白发生反应的单克隆抗体
DOI：
10.1002/jcb.240320307
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发表时间：
1986
影响因子：
4
通讯作者：
H. Wolfe
中科院分区：
文献类型：
--
作者：
W. Carney;Hamer Pj;D. Petit;H. Rabin;G. Cooper;M. Lefebvre;H. Wolfe
活化的ras转化基因已在多种肿瘤中被描述，并编码21，000-道尔顿（p21）蛋白，在位置12、13和61处具有氨基酸取代。在这份报告中，我们描述了一种单克隆抗体命名为DWP的反应。特别是在12位含有缬氨酸的合成十二肽，在较小程度上在12位含有半胱氨酸的肽，而不是在相同位置含有甘氨酸、精氨酸、丝氨酸、天冬氨酸、谷氨酸或丙氨酸的肽。Western印迹和免疫过氧化物酶研究表明，DWP特异性反应与激活rasH或rasK蛋白在NIH细胞转化的DNA从人癌细胞编码缬氨酸在位置12。DWP不与编码甘氨酸的正常p21在位置12处反应，也不与编码天冬氨酸、谷氨酸、精氨酸、丝氨酸或半胱氨酸的激活p21在位置12处反应。对人肿瘤细胞系的调查表明，DWP与人膀胱癌细胞系T24反应，但不与先前显示的在位置12或61处含有其他激活突变的人肿瘤细胞系反应。DWP和可能与ras蛋白12或61位的改变特异性反应的其他抗体在确定人类恶性肿瘤中活化ras蛋白的存在和频率方面可能是有价值的。
Activated ras transforming genes have been described in a variety of neoplasms and encode 21,000‐Dalton (p21) proteins with amino acid substitutions at positions 12, 13, and 61. In this report we describe a monoclonal antibody designated DWP that reacts. Specifically with synthetic dodecapeptides containing valine at position 12, to a lesser extent with peptides containing cysteine at position 12 and not with peptides containing glycine, arginine, serine, aspartic acid, glutamic acid or alanine at the same position. Western blot and immunoperoxidase studies showed that DWP specifically reacts with activated rasH or rasK proteins in NIH cells transformed by DNA from the human carcinoma cells that encode valine at position 12. DWP did not react with normal p21s encoding glycine at position 12, nor with activated p21s encoding aspartic acid, glutamic acid, arginine, serine, or cysteine at position 12. A survey of human tumor cell lines demonstrated that DWP reacted with the human bladder carcinoma cell line T24 but not with human tumor cell lines previously shown td contain other activating mutations at positions 12 or 61. DWP and perhaps additional antibodies that specifically react with alterations at positions 12 or 61 of the ras protein may be valuable in determining the presence and frequency of activated ras proteins in human malignancy.