Führer Sebastian, Ahammer Linda, Ausserbichler Angela, Scheffzek Klaus, Dunzendorfer-Matt Theresia, Tollinger Martin
Center for Molecular Biosciences Innsbruck (CMBI), Institute of Organic Chemistry, University of Innsbruck, Innrain 80/82, 6020, Innsbruck, Austria.
Division of Biological Chemistry, Biocenter, Medical University of Innsbruck, Innrain 80/82, 6020, Innsbruck, Austria.
Biomol NMR Assign. 2017 Oct;11(2):305-308. doi: 10.1007/s12104-017-9768-1. Epub 2017 Aug 22.
Neurofibromin and Sprouty-related EVH1 domain-containing protein 1 (Spred1) both act as negative regulators of the mitogen-activated protein kinase pathway and are associated with the rare diseases Neurofibromatosis type 1 and Legius syndrome, respectively. Spred1 recruits the major GTPase activating protein (GAP) neurofibromin from the cytosol to the membrane in order to inactivate the small G protein Ras. These functions are dependent on the N-terminal EVH1 domain and the C-terminal Sprouty domain of Spred1 whereas the former specifically recognizes the GAP related domain of neurofibromin and the latter is responsible for membrane targeting. Within the GAP domain, Spred1 binding depends on the GAPex portion which is dispensable for Ras inactivation. In a first step towards the characterization of the Neurofibromin Spred1 interface in solution we assigned backbone and side chain H, C, and N chemical shifts of the Spred1 derived EVH1 domain. Our chemical shift data analysis indicate seven consecutive β-strands followed by a C-terminal α-helix which is in agreement with the previously reported crystal structure of Spred1(EVH1). Our data provide a framework for further analysis of the function of patient-derived mutations associated with rare diseases.
神经纤维瘤蛋白和含Sprouty相关EVH1结构域蛋白1（Spred1）均作为丝裂原活化蛋白激酶途径的负调节因子，分别与罕见疾病1型神经纤维瘤病和Legius综合征相关。Spred1将主要的GTP酶激活蛋白（GAP）神经纤维瘤蛋白从细胞质募集到细胞膜，从而使小G蛋白Ras失活。这些功能依赖于Spred1的N端EVH1结构域和C端Sprouty结构域，前者特异性识别神经纤维瘤蛋白的GAP相关结构域，后者负责膜靶向。在GAP结构域内，Spred1的结合取决于对Ras失活可有可无的GAPex部分。作为在溶液中表征神经纤维瘤蛋白Spred1界面的第一步，我们确定了Spred1衍生的EVH1结构域的主链和侧链H、C和N化学位移。我们的化学位移数据分析表明有七个连续的β链，接着是一个C端α螺旋，这与先前报道的Spred1（EVH1）晶体结构一致。我们的数据为进一步分析与罕见疾病相关的患者来源突变的功能提供了框架。
