Monoclonal antibody specific for an activated RAS protein.
Monoclonal antibody specific for an activated RAS protein.
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针对活化 RAS 蛋白的单克隆抗体。
DOI：
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发表时间：
1986
影响因子：
11.1
通讯作者：
A. Tischler
中科院分区：
文献类型：
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作者：
W. Carney;D. Petit;P. Hamer;C. Der;T. Finkel;G. Cooper;M. Lefebvre;H. Mobtaker;R. DeLellis;A. Tischler
已在10-20%的人类肿瘤中检测到编码在位置12、13或61处具有氨基酸取代的蛋白质（p21）的活化RAS转化基因。本报告描述了针对对应于突变RAS基因的氨基酸5-16的合成肽产生的单克隆抗体（DWP），所述突变RAS基因在位置12处编码瓦尔而不是Gly。DWP在竞争测定中与在位置12处含有瓦尔或Cys的肽反应，但不与在位置12处含有Gly、Arg、Ser、Ala、Asp或Glu的肽反应。转化的NIH细胞和人癌细胞系的免疫印迹分析显示，DWP与在位置12处含有瓦尔的活化RAS蛋白特异性反应，而不与正常p21或通过在位置12和61处的其它氨基酸取代活化的p21反应。免疫组织化学研究表明，DWP标记的转化NIH细胞和人癌细胞含有p21，其12位为瓦尔或Cys，但不含正常或其他活化的p21。与人癌细胞系所见的特异性相反，福尔马林固定的原发性癌标本的分析表明，阳性免疫过氧化物酶染色与DWP不一定与免疫印迹和转染测定活化RAS蛋白的存在相关。然而，免疫组织化学研究表明，DWP优先结合人类癌细胞。
Activated RAS transforming genes that encode proteins (p21s) with amino acid substitutions at positions 12, 13, or 61 have been detected in 10-20% of human neoplasms. This report describes a monoclonal antibody (DWP) raised against a synthetic peptide corresponding to amino acids 5-16 of a mutated RAS gene encoding Val instead of Gly at position 12. DWP reacted in competition assays with peptides containing Val or Cys at position 12, but did not react with peptides containing Gly, Arg, Ser, Ala, Asp, or Glu at position 12. Immunoblot analysis of transformed NIH cells and human carcinoma cell lines showed that DWP reacts specifically with activated RAS proteins containing Val at position 12 and not with normal p21s or p21s activated by other amino acid substitutions at positions 12 and 61. Immunohistochemical studies showed that DWP-labeled transformed NIH cells and human carcinoma cells contained p21s with either Val or Cys at position 12 but not normal or other activated p21s. In contrast to the specificity seen with human carcinoma cell lines, analysis of formalin-fixed, primary carcinoma specimens indicated that positive immunoperoxidase staining with DWP did not necessarily correlate with immunoblot and transfection assays for the presence of activated RAS proteins. Immunohistochemical studies did show, however, that DWP preferentially binds human carcinoma cells.