Mutagenesis of the H-ras p21 at glycine-60 residue disrupts GTP-induced conformational change.
Mutagenesis of the H-ras p21 at glycine-60 residue disrupts GTP-induced conformational change.
复制标题
H-ras p21 在甘氨酸 60 残基处的诱变破坏了 GTP 诱导的构象变化。
DOI：
10.1021/bi00010a040
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发表时间：
1995
期刊：
影响因子：
2.9
通讯作者：
Hwang,YW
中科院分区：
文献类型：
--
作者：
Sung,YJ;Carter,M;Zhong,JM;Hwang,YW
1994年12月21日收到的修订稿®摘要；Gly-60是vH-ras DXXG结构域的保守甘氨酸，通过定点诱变检测其功能。结果表明，G60A （gy -60 to Ala substitution）突变对H-ras与鸟嘌呤核苷酸的相互作用影响不大，但完全消除了vH-ras的生物活性。G60A突变对H-ras与SDC25C（鸟嘌呤核苷酸交换因子）和GAP的相互作用也没有影响。然而，G60A突变确实降低了H-ras结合Raf的能力。GTP诱导H-ras与荧光染料8-苯胺- 1 -萘磺酸组成的配合物的荧光发射增强。这种增强被G60A突变阻断。基于这些观察结果，我们提出G60A突变破坏了gtp诱导的H-ras构象变化，这是H-ras活性所必需的过程。
Revised Manuscript Received December 21, 1994® abstract; The function of Gly-60, the conserved glycine in the DXXG domain of vH-ras, was examined by site-directed mutagenesis. It was found that while the G60A (Gly-60 to Ala substitution) mutation has little effect on the interaction of H-ras with guanine nucleotides, it completely abolishesthe biological activity of vH-ras. The G60A mutation also exerts little effect on the interaction of H-ras with SDC25C (a guanine nucleotide exchange factor) and GAP. However, the G60A mutation does lower the ability of H-ras to bind Raf. GTP induces an enhancement of fluorescence emission in complexes consisting of H-ras and the fluorescent dye 8-anilino-l-naphthalenesulfonic acid. This enhancement is blocked by the G60A mutation. On the basis of these observations, we propose that the GTP-induced conformational change of H-ras, a process required for H-ras activities, is impaired by the G60A mutation.