DESIGN OF SSPB VARIANTS FOR PROBING SSPB FUNCTION

用于探测 SSPB 功能的 SSPB 变体的设计

• 批准号: 7721200
• 负责人: Robert T Sauer
• 金额: $ 0.7万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-05-15 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7721200
• 关键词: Adaptor Signaling Protein; Alanine; Binding; Cleaved cell; Computer Retrieval of Information on Scientific Projects Database; Cysteine; Engineering; Funding; Glutamine; Grant; Institution; Modeling; Mutate; Process; Property; Proteins; Reagent; Research; Research Personnel; Resources; Source; Testing; United States National Institutes of Health; Variant; crosslink; design; dimer; monomer; novel; protein function; research study; tool

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Protein design is a useful tool for creating novel reagents for probing the function of proteins. Dan Bolon of the Sauer lab has created several designed variants of adaptor protein SspB that he has used to test models of SspB function. The features of SspB related to function that were subjected to the design process were the dimer interface (SspB forms a symmetric dimer), the substrate binding cleft and the c-terminal extensions from the substrate binding domain that have been shown to interact directly with ClpX. In one design, cysteines were engineered into SspB and its substrate, the ssrA tag, in order to crosslink the adaptor and its substrate. In another design, alanine 74 of SspB was mutated to glutamine. This design was intended to interfere with substrate binding. The third design experiment re-engineered the SspB dimer interface so that it would not be symmetric, allowing formation of heterodimers between monomers with different substrate- and/or ClpX-binding properties.

该子项目是利用该技术的众多研究子项目之一 资源由 NIH/NCRR 资助的中心拨款提供。子项目及 研究者 (PI) 可能已从 NIH 的另一个来源获得主要资金， 因此可以在其他 CRISP 条目中表示。列出的机构是 对于中心来说，它不一定是研究者的机构。 蛋白质设计是创建用于探测蛋白质功能的新型试剂的有用工具。 Sauer 实验室的 Dan Bolon 创建了几种接头蛋白 SspB 的设计变体，他用它们来测试 SspB 功能模型。设计过程中与功能相关的 SspB 特征是二聚体界面（SspB 形成对称二聚体）、底物结合裂口和底物结合结构域的 C 端延伸（已显示可直接与 ClpX 相互作用） 。在一种设计中，半胱氨酸被工程化到 SspB 及其底物 ssrA 标签中，以便交联接头及其底物。 在另一种设计中，SspB 的丙氨酸 74 突变为谷氨酰胺。 该设计旨在干扰底物结合。 第三个设计实验重新设计了 SspB 二聚体界面，使其不对称，从而允许具有不同底物和/或 ClpX 结合特性的单体之间形成异二聚体。