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本文引用的文献

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Confirmation of the arginine-finger hypothesis for the GAP-stimulated GTP-hydrolysis reaction of Ras.对Ras的GAP刺激的GTP水解反应的精氨酸指假说的证实。
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Equilibrium and kinetic study of the conformational transition toward the active state of p21Ha-ras, induced by the binding of BeF3- to the GDP-bound state, in the absence of GTPase-activating proteins.
J Biol Chem. 1997 Sep 12;272(37):23138-43. doi: 10.1074/jbc.272.37.23138.
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Calculation of pathways for the conformational transition between the GTP- and GDP-bound states of the Ha-ras-p21 protein: calculations with explicit solvent simulations and comparison with calculations in vacuum.Ha-ras-p21蛋白结合GTP和GDP状态之间构象转变途径的计算：显式溶剂模拟计算及与真空计算的比较
Proteins. 1997 Jul;28(3):434-51.
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Formation of a transition-state analog of the Ras GTPase reaction by Ras-GDP, tetrafluoroaluminate, and GTPase-activating proteins.由Ras-GDP、四氟铝酸盐和GTP酶激活蛋白形成Ras GTP酶反应的过渡态类似物。
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Function and regulation of ras.Ras的功能与调控
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Mechanism of GTP hydrolysis by p21N-ras catalyzed by GAP: studies with a fluorescent GTP analogue.由GAP催化的p21N-ras对GTP的水解机制：使用荧光GTP类似物的研究
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Long PCR.长片段聚合酶链反应
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Activators and effectors of ras p21 proteins.Ras p21蛋白的激活剂和效应器。
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Solution structure and dynamics of ras p21.GDP determined by heteronuclear three- and four-dimensional NMR spectroscopy.通过异核三维和四维核磁共振光谱法测定的ras p21.GDP的溶液结构与动力学
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Ras蛋白激活反应途径中效应环铰链区的特征。三氟化铍与Ha-ras-p21的V29G和I36G突变体结合的动力学。

Characterization of the hinges of the effector loop in the reaction pathway of the activation of ras-proteins. Kinetics of binding of beryllium trifluoride to V29G and I36G mutants of Ha-ras-p21.

作者信息

Kuppens S, Díaz J F, Engelborghs Y

机构信息

Laboratory of Biomolecular Dynamics, Katholieke Universiteit Leuven, Heverlee, Belgium.

出版信息

Protein Sci. 1999 Sep;8(9):1860-6. doi: 10.1110/ps.8.9.1860.

DOI:10.1110/ps.8.9.1860
PMID:10493587
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC2144410/
Abstract

This work experimentally confirms the pathway of activation of Ha-ras-p21, which was calculated by the method of Targeted Molecular Dynamics (TMD) (Díaz JF, Wroblowski B, Schlitter J, Engelborghs Y, 1997a, Proteins Struct Funct Genet 28:434-451). The process can be studied experimentally by analyzing the binding of BeF3- to the GDP complex of the active fluorescent mutant Y32W (Díaz JF, Sillen A, Engelborghs Y, 1997b, J Biol Chem 227:23138-23143). Two mutants, V29G and 136G, have been constructed at both sides of the effector loop of the active fluorescent mutant. This was done to check the proposed reaction pathway and to provide further insight into the mechanism of the activation of ras proteins. Both mutations accelerate the conformational isomerization with two orders of magnitude, demonstrating convincingly the role of these residues as hinges of the effector loop in one or more of the transitions of the conformational change. These results provide experimental support to the pathway calculated by TMD analysis.

摘要

这项工作通过实验证实了Ha-ras-p21的激活途径，该途径是通过靶向分子动力学（TMD）方法计算得出的（迪亚兹JF、沃罗布罗斯基B、施利特J、恩格尔博格斯Y，1997a，《蛋白质结构、功能与遗传学》28:434 - 451）。可以通过分析BeF3-与活性荧光突变体Y32W的GDP复合物的结合来对该过程进行实验研究（迪亚兹JF、西伦A、恩格尔博格斯Y，1997b，《生物化学杂志》227:23138 - 23143）。在活性荧光突变体的效应环两侧构建了两个突变体V29G和I36G。这样做是为了检验所提出的反应途径，并进一步深入了解ras蛋白的激活机制。这两个突变均使构象异构化加速了两个数量级，令人信服地证明了这些残基在构象变化的一个或多个转变中作为效应环铰链的作用。这些结果为通过TMD分析计算出的途径提供了实验支持。