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.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 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 Lab的Dan Bolon创建了几种设计的适配器蛋白SSPB的变体，他用来测试SSPB功能的模型。与函数相关的SSPB的特征是遵循设计过程的功能，是二聚体界面（SSPB形成对称二聚体），底物结合裂口和来自底物结合结构域的C末端扩展，这些域已显示出直接与Clpx相互作用。在一个设计中，半胱氨酸被设计到SSPB及其基板SSRA标签中，以交叉链接适配器及其底物。 在另一种设计中，将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引入了SSPB二聚体界面，因此它不会对称，从而允许在具有不同基板和/或CLPX结合属性的单体之间形成异二聚体。