MOLECULAR-CLONING OF A GTPASE ACTIVATING PROTEIN-SPECIFIC FOR THE KREV-1 PROTEIN P21RAP1
MOLECULAR-CLONING OF A GTPASE ACTIVATING PROTEIN-SPECIFIC FOR THE KREV-1 PROTEIN P21RAP1
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DOI：
10.1016/0092-8674(91)90555-d
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发表时间：
1991-06-14
期刊：
影响因子：
64.5
通讯作者：
POLAKIS, P
中科院分区：
文献类型：
--
作者：
RUBINFELD, B;MUNEMITSU, S;POLAKIS, P
rap1/Krev-1基因编码ras相关蛋白，抑制ras癌基因的转化。我们从牛脑中纯化了rap1/Krev-1基因产物特异性的88 kd GTPase激活蛋白（GAP）。根据该蛋白的部分氨基酸序列，从人脑文库中分离出3.3 kb的cDNA。cDNA在昆虫Sf9细胞中的表达导致85-95 kd rap1GAP的高水平产生，该gap特异性地刺激了p2l（rap1）的GTPase活性。完整的氨基酸序列与任何已知的蛋白质序列都不同源，包括p2l（ras）特有的gap。Northern和Western blotting分析表明，rapgap不是普遍表达的，在胎儿组织和某些肿瘤细胞系中表达最多，特别是Wilms肾肿瘤SK-NEP-1和黑色素瘤SK-MEL-3细胞系。
The rap1/Krev-1 gene encodes a ras-related protein that suppresses transformation by ras oncogenes. We have purified an 88 kd GTPase activating protein (GAP), specific for the rap1/Krev-1 gene product, from bovine brain. Based on partial amino acid sequences obtained from this protein, a 3.3 kb cDNA was isolated f rom a human brain library. Expression of the cDNA in insect Sf9 cells resulted in high level production of an 85-95 kd rap1GAP that specifically stimulated the GTPase activity of p2l(rap1). The complete deduced amino acid sequence is not homologous to any known protein sequences, including GAPs specific for p2l(ras). Northern and Western blotting analysis indicate that raplGAP is not ubiquitously expressed and appears most abundant in fetal tissues and certain tumor cell lines, particularly the Wilms' kidney tumor, SK-NEP-1, and the melanoma, SK-MEL-3, cell lines.