MAMMALIAN RAS INTERACTS DIRECTLY WITH THE SERINE THREONINE KINASE RAF
MAMMALIAN RAS INTERACTS DIRECTLY WITH THE SERINE THREONINE KINASE RAF
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DOI：
10.1016/0092-8674(93)90307-c
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发表时间：
1993-07-16
期刊：
影响因子：
64.5
通讯作者：
COOPER, JA
中科院分区：
文献类型：
--
作者：
VOJTEK, AB;HOLLENBERG, SM;COOPER, JA
我们已经确定了蛋白质相互作用的H-Ras使用双杂交系统筛选的小鼠cDNA文库。大约50%的克隆鉴定了c-Raf和A-Raf丝氨酸/苏氨酸激酶的编码部分。这些克隆之间的重叠将Raf的N-末端的保守的81个残基区域定义为Ras相互作用区域。我们发现，Raf与野生型和激活的Ras相互作用，但不与Ras的效应域突变体或显性干扰Ras突变体。使用纯化的细菌表达的融合蛋白，我们表明，此外，Ras和Raf的N-末端区域直接在体外关联，这种相互作用是依赖于GTP结合Ras。
We have identified proteins that interact with H-Ras using a two hybrid system screen of a mouse cDNA library. Approximately 50% of the clones identified encoded portions of the c-Raf and A-Raf serine/threonine kinases. Overlaps among these clones define a conserved 81 residue region of the N-terminus of Raf as the Ras interaction region. We show that Raf interacts with wild-type and activated Ras, but not with an effector domain mutant of Ras or with a dominant-interfering Ras mutant. Using purified bacterially expressed fusion proteins, we show, furthermore, that Ras and the N-terminal region of Raf associate directly in vitro and that this interaction is dependent on GTP bound to Ras.