SMALL ANGLE X-RAY SCATTERING OF F FACTOR TRAI AND ITS DOMAINS

F因子TRAI及其域的小角X射线散射

• 批准号: 8363565
• 负责人: JOEL F SCHILDBACH
• 金额: $ 1.04万
• 依托单位: CORNELL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-07-01 至 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8363565
• 关键词: C-terminal; Cleaved cell; DNA Binding; Data; F Factor; Funding; Grant; N-terminal; National Center for Research Resources; Plasmids; Principal Investigator; Process; Proteins; Relative (related person); Research; Research Infrastructure; Resources; Roentgen Rays; Shapes; Source; Structure; United States National Institutes of Health; cost; helicase; polypeptide; research study

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. Primary support for the subproject and the subproject's principal investigator may have been provided by other sources, including other NIH sources. The Total Cost listed for the subproject likely represents the estimated amount of Center infrastructure utilized by the subproject, not direct funding provided by the NCRR grant to the subproject or subproject staff. TraI, a protein from the conjugative plasmid F factor, is essential for the plasmid transfer process. The protein includes a relaxase activity that cleaves one plasmid strand prior to transfer, and a helicase activity ---- both of which must be in one polypeptide chain for efficient conjugative transfer. We have determined the crystal structure of the relaxase domain (the N-terminal 36 kDa of the 192 kDa protein), and are near solving the 3D NMR structure of a second 20 kDa region. The C-terminal domain has been solved, and we are pursuing structures of other domains. We want to collect SAXS data to determine the shape of the intact protein and various fragments, and attempt to fit the regions of known structure into the resulting shapes. The experiments will provide information on the orientation of the domains relative to each other and whether the orientation of the domains changes upon DNA binding.

该子项目是利用资源的众多研究子项目之一 由 NIH/NCRR 资助的中心拨款提供。子项目的主要支持 并且子项目的主要研究者可能是由其他来源提供的， 包括其他 NIH 来源。 子项目可能列出的总成本 代表子项目使用的中心基础设施的估计数量， NCRR 赠款不直接向子项目或子项目工作人员提供资金。 TraI 是一种来自接合质粒 F 因子的蛋白质，对于质粒转移过程至关重要。 该蛋白质包括在转移前切割一条质粒链的松弛酶活性和解旋酶活性——这两者必须位于一条多肽链中才能有效的接合转移。 我们已经确定了松弛酶结构域（192 kDa 蛋白质的 N 端 36 kDa）的晶体结构，并且即将解决第二个 20 kDa 区域的 3D NMR 结构。 C端结构域已经解决，我们正在研究其他结构域的结构。 我们想要收集 SAXS 数据来确定完整蛋白质和各种片段的形状，并尝试将已知结构的区域拟合到最终的形状中。 实验将提供有关结构域相对于彼此的方向以及结构域的方向是否在 DNA 结合时发生变化的信息。