磷酸酪氨酸配体与Grb2衔接蛋白的SH2结构域结合后，该衔接蛋白结构动力学的变化。

Changes in structural dynamics of the Grb2 adaptor protein upon binding of phosphotyrosine ligand to its SH2 domain.

作者信息

de Mol Nico J, Catalina M Isabel, Fischer Marcel J E, Broutin Isabelle, Maier Claudia S, Heck Albert J R

机构信息

Department of Medicinal Chemistry, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, P.O. Box 80082, Sorbonnelaan 16, 3508 TB Utrecht, The Netherlands.

出版信息

Biochim Biophys Acta. 2004 Jul 1;1700(1):53-64. doi: 10.1016/j.bbapap.2004.03.016.

DOI:10.1016/j.bbapap.2004.03.016

PMID:15210125

Abstract

Growth factor receptor-bound protein 2 (Grb2) is an extensively studied adaptor protein involved in cell signaling. Grb2 is a highly flexible protein composed of a single SH2 domain flanked by two SH3 domains. Here we report on the structural dynamic effects upon interaction of a phosphopeptide ligand derived from the recognition sequence of the Shc adaptor protein with (i) the isolated SH2 domain of Grb2 (Grb2 SH2) and (ii) the full-length Grb2 protein. From kinetic studies using surface plasmon resonance, it was deduced that a conformation change occurred in the SH2 protein as well as the full-length Grb2 after binding. Measurements of hydrogen/deuterium exchange (HDX) in the isolated SH2 domain and full-length Grb2 protein as monitored by electrospray mass spectrometry, showed that binding reduces the overall flexibility of the proteins, possibly via slightly different mechanisms for the single SH2 domain and the full-length Grb2 protein.

摘要

生长因子受体结合蛋白2（Grb2）是一种参与细胞信号传导且被广泛研究的衔接蛋白。Grb2是一种高度灵活的蛋白质，由一个位于两个SH3结构域之间的单一SH2结构域组成。在此，我们报告了源自Shc衔接蛋白识别序列的磷酸肽配体与（i）Grb2的分离SH2结构域（Grb2 SH2）和（ii）全长Grb2蛋白相互作用时的结构动力学效应。通过表面等离子体共振进行的动力学研究推断，结合后SH2蛋白以及全长Grb2中均发生了构象变化。用电喷雾质谱监测分离的SH2结构域和全长Grb2蛋白中的氢/氘交换（HDX）测量结果表明，结合降低了蛋白质的整体灵活性，可能是通过单一SH2结构域和全长Grb2蛋白的略有不同的机制实现的。