SAXS STUDIES OF BTB PROTEIN COMPLEXES

BTB 蛋白质复合物的 SAXS 研究

• 批准号: 7954932
• 负责人: GILBERT G PRIVE
• 金额: $ 0.65万
• 依托单位: ILLINOIS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-01 至 2009-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7954932
• 关键词: BTB/POZ Domain; Back; Binding; Biophysics; C-terminal; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA; DNA Binding; Data; Disease; Funding; Grant; Housing; Human; Institution; Length; N-terminal; Proteins; Research; Research Personnel; Resources; Solutions; Source; Structure; Ubiquitination; United States National Institutes of Health; Zinc Fingers; base; protein complex; transcription factor; 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. There are 43 BTB-ZF and 49 BTB-BACK-Kelch (BBK) proteins in humans (Stogios et al 2005). The BTB-ZF proteins have a single N-terminal BTB domain, an intrinsically disordered middle region and a set of C terminal zinc-finger (ZF) domains. Similarly, the BBK proteins have a single N-terminal BTB domain, a central BACK domain (Stogios and Priv¿ 2004) and C-terminal Kelch repeats. In general, all BTB-ZF proteins are transcription factors, and the BBK proteins are substrate binding adaptors in Cullin3-based E3 ubiquitin ligase complexes. BTB-ZF proteins: We have solved a total of 9 crystal structures of BTB domains from BTB-ZF proteins (Ghetu et al 2008; Stogios et al 2007, Ahmad et al 2003, etc). These are either homodimers or hexamers, implying that the transcription factors are either bivalent or hexavalent. We are now moving towards studying the 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. BBK proteins are elongated dimeric and/or tetrameric proteins that interact with Cul3 at the BTB end and substrate at the Kelch end, resulting in the ubiquitination of substrate in Cul3-based SCF-like E3 ligase complexes (Pintard et al 2004). 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 2:2 and 4:4 BBK/Cul3 complexes.

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。子项目和 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。 人类中有 43 个 BTB-ZF 和 49 个 BTB-BACK-Kelch (BBK) 蛋白 (Stogios et al 2005)。 BTB-ZF 蛋白具有单个 N 端 BTB 结构域、本质上无序的中间区域和一组 C 端锌指 (ZF) 结构域。 类似地，BBK 蛋白具有单个 N 端 BTB 结构域、中央 BACK 结构域（Stogios 和 Priv' 2004）和 C 端 Kelch 重复序列。一般来说，所有 BTB-ZF 蛋白都是转录因子，BBK 蛋白是基于 Cullin3 的 E3 泛素连接酶复合物中的底物结合接头。 BTB-ZF 蛋白：我们已经从 BTB-ZF 蛋白中解析出总共 9 个 BTB 结构域的晶体结构（Ghetu 等人 2008；Stogios 等人 2007；Ahmad 等人 2003 等）。这些是同二聚体或六聚体，这意味着转录因子是二价或六价的。 我们现在正着手通过 SAXS 研究全长蛋白质，以便了解全长蛋白质内 DNA 结合 ZF 结构域的拓扑排列，可能与 DNA 片段复合。 BBK 蛋白是细长的二聚体和/或四聚体蛋白，可与 BTB 端的 Cul3 和 Kelch 端的底物相互作用，导致基于 Cul3 的 SCF 样 E3 连接酶复合物中的底物泛素化 (Pintard et al 2004)。 E3 连接酶复合物已被广泛研究，但人们对这些复合物的寡聚状态问题知之甚少。 BBK 蛋白通过其 BTB 结构域寡聚，我们的内部解决方案数据显示 2:2 和 4:4 BBK/Cul3 复合物。