SAXS STUDIES OF BTB PROTEIN COMPLEXES

BTB 蛋白质复合物的 SAXS 研究

• 批准号: 8168633
• 负责人: GILBERT G PRIVE
• 金额: $ 0.54万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-01-01 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8168633
• 关键词: Adaptor Signaling Protein; Affect; Architecture; BTB/POZ Domain; Back; Binding; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Binding; Data; Funding; Gel Chromatography; Grant; Housing; Human; Institution; Length; Proteins; Research; Research Personnel; Resources; Series; Solutions; Source; Structure; United States National Institutes of Health; ZNF145 gene; Zinc Fingers; dimer; interest; protein complex; protein protein interaction; transcription factor; ubiquitin ligase; ubiquitin-protein ligase

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. BTB domains are protein-protein interaction motifs can that form homomeric and heteromeric complexes. There are 43 BTB-Zinc Finger and 49 BTB-BACK-Kelch (BBK) proteins in humans (Stogios et al 2005). The BTB-ZF proteins are generally transcription factors, and the BBK proteins are generally adaptor proteins in cullin ubiquitin ligase E3 complexes. In both BTB-ZF and BBK proteins, the BTB domains self associate into higher oligomeric species. We are interested in studying how BTB domains affect the architectures of transcription factors and ubiquitin ligase complexes. 1) BTB-ZF proteins. We have solved a total of 9 crystal structures of BTB domains from BTB-ZF proteins with data collected at the APS (Ghetu et al 2008; Stogios et al 2007, Ahmad et al 2003, etc). These BTB domains are either homodimers or hexamers, implying that there are either two or six DNA binding motifs in the full length transcription factors. We plan to study a series of full-length proteins by SAXS in order to understand the topological arrangement of the DNA binding ZF domains within the full-length proteins, possibly in complex with DNA fragments. We have expressed nearly full-length PLZF and it is a monodisperse dimer by gel filtration and DLS. 2) BBK proteins. E3 ligase complexes are widely studied, but the issue of the oligomeric state of these complexes is poorly understood. BBK proteins oligomerize via their BTB domains, and we have in-house solution data showing both dimeric (2:2) and tetrameric (4:4) BBK/cullin complexes. We plan to collect SAXS data on a number of BBK/cullin complexes that have different stoicheometries in order to understand how BBK proteins orient their bound substrates in E3 complexes.

这个子项目是许多研究子项目中的一个 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得了主要资金， 因此可以在其他CRISP条目中表示。所列机构为 研究中心，而研究中心不一定是研究者所在的机构。 BTB结构域是蛋白质-蛋白质相互作用基序，可以形成同聚和异聚复合物。 人体中有43种BTB-锌指蛋白和49种BTB-BACK-Kelch（BBK）蛋白（Stogios et al 2005）。 BTB-ZF蛋白通常是转录因子，BBK蛋白通常是cullin泛素连接酶E3复合物中的衔接蛋白。在BTB-ZF和BBK蛋白中，BTB结构域自缔合成更高的寡聚物种类。 我们有兴趣研究BTB结构域如何影响转录因子和泛素连接酶复合物的结构。1)BTB-ZF蛋白。我们已经用在APS收集的数据解析了来自BTB-ZF蛋白的BTB结构域的总共9种晶体结构（Ghetu等2008; Stogios等2007，Ahmad等2003等）。这些BTB结构域是同源二聚体或六聚体，这意味着在全长转录因子中存在两个或六个DNA结合基序。 我们计划通过SAXS研究一系列全长蛋白质，以了解全长蛋白质中DNA结合ZF结构域的拓扑排列，可能与DNA片段复合。 我们表达了几乎全长的PLZF，通过凝胶过滤和DLS，它是一个单分散的二聚体。2)BBK蛋白。 E3连接酶复合物被广泛研究，但这些复合物的寡聚状态的问题知之甚少。 BBK蛋白通过其BTB结构域寡聚化，并且我们有内部溶液数据显示二聚体（2：2）和四聚体（4：4）BBK/cullin复合物。我们计划收集SAXS数据的BBK/cullin复合物，有不同的化学计量，以了解如何BBK蛋白定向其结合底物在E3复合物。