COMPLEXES OF RAS.GTP WITH RAF-1 AND MITOGEN-ACTIVATED PROTEIN-KINASE KINASE
COMPLEXES OF RAS.GTP WITH RAF-1 AND MITOGEN-ACTIVATED PROTEIN-KINASE KINASE
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DOI：
10.1126/science.8503013
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发表时间：
1993-06-11
期刊：
影响因子：
56.9
通讯作者：
WOLFMAN, A
中科院分区：
文献类型：
--
作者：
MOODIE, SA;WILLUMSEN, BM;WOLFMAN, A
鸟苷三磷酸（GTP）结合蛋白Ras在调节生长和分化中起作用;然而，对引起其生物活性的蛋白质相互作用知之甚少。将野生型Ras或Ras的突变形式共价连接到不溶性基质上，然后用于检查信号蛋白与Ras的相互作用。Ras的形式激活突变（Gly 12 Val）或GTP类似物，鸟苷酰亚氨基二磷酸（GMP-PNP）的结合，特别是与Raf-1相互作用，而效应域突变体，Ile 36 Ala，未能与Raf-1相互作用。丝裂原活化蛋白激酶（MAP激酶）活性仅与活化形式的Ras相关。通过直接测定证实了活化的Ras与活化的MAP激酶激酶（MAPK）的特异性相互作用。因此，含有MAPKK活性和Raf-1蛋白的复合物的形成依赖于Ras的活性。
The guanosine triphosphate (GTP)-binding protein Ras functions in regulating growth and differentiation; however, little is known about the protein interactions that bring about its biological activity. Wild-type Ras or mutant forms of Ras were covalently attached to an insoluble matrix and then used to examine the interaction of signaling proteins with Ras. Forms of Ras activated either by mutation (Gly12Val) or by binding of the GTP analog, guanylyl-imidodiphosphate (GMP-PNP) interacted specifically with Raf-1 whereas an effector domain mutant, Ile36Ala, failed to interact with Raf-1. Mitogen-activated protein kinase (MAP kinase) activity was only associated with activated forms of Ras. The specific interaction of activated Ras with active MAP kinase kinase (MAPK) was confirmed by direct assays. Thus the forming of complexes containing MAPKK activity and Raf-1 protein are dependent upon the activity of Ras.