STRUCTURAL STUDY OF MURINE GAMMA-CRYSTALLIN BY COMBINED USE OF SAXS AND WEAK ALI

结合使用 SAXS 和弱 ALI 的鼠γ-晶状体蛋白结构研究

• 批准号: 7370523
• 负责人: JILL TREWHELLA
• 金额: $ 0.49万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-03-01 至 2007-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7370523
• 关键词: STRUCTURAL; STUDY; MURINE; GAMMA; CRYSTALLIN

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. Using a procedure termed molecular fragment replacement (MFR) it has become possible to rapidly solve protein structures by NMR using only residual dipolar couplings (RDCs) and chemical shifts, even in the complete absence of NOE or dihedral information. Application of this technology to murine g-crystallin yields structures for its two globular domains that are in good agreement with x-ray structures of other previously studied g-crystallins. Although the dipolar coupling information provides very tight restraints for the relative orientation of the two g-crystallin domains, the NMR data lack translational information. Simulations indicate that the SAXS data will yield this critical information. High quality experimental SAXS data on murine g-crystallin is needed to demonstrate that the combination of SAXS and weak alignment NMR provides a new and powerful tool for rapidly determining structures of macromolecules and their complexes. With the structure of globular domains frequently known in advance, and their relative orientation well determined by the dipolar couplings obtained from weak alignment NMR, high signal-to-noise SAXS data (obtained at a S/N ratio of at least ca 3:1 at 0.5/¿¿, provides the translational information needed to rapidly solve the structure of multi-module proteins, and protein-protein and protein-nucleic acid complexes. If successful on this system, this approach could be applied to a wide variety of modular or multidomain proteins in solution, providing insights into domain interactions under a variety of conditions and functional states.

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。使用称为分子片段置换（MFR）的程序，即使在完全不存在NOE或二面角信息的情况下，也可以通过NMR仅使用残余偶极偶联（RDC）和化学位移快速解析蛋白质结构。这种技术的应用，以鼠g-晶体蛋白产生的结构，其两个球状域是在良好的协议与其他先前研究的g-晶体蛋白的X-射线结构。 虽然偶极耦合信息提供了非常严格的限制两个g-晶体蛋白域的相对取向，NMR数据缺乏翻译信息。模拟表明，SAXS数据将产生这一关键信息。高质量的实验SAXS数据对小鼠g-晶状体蛋白是必要的，以证明SAXS和弱排列NMR的组合提供了一个新的和强大的工具，用于快速确定大分子及其复合物的结构。 由于球状结构域的结构通常是预先已知的，并且它们的相对取向通过从弱排列NMR获得的偶极偶联很好地确定，高信噪比SAXS数据（在0.5/μ S下以至少约3：1的S/N比获得）提供了快速解析多模块蛋白质、蛋白质-蛋白质和蛋白质-核酸复合物的结构所需的翻译信息。如果在这个系统上成功，这种方法可以应用于各种各样的模块化或多结构域蛋白质的解决方案，在各种条件和功能状态下提供域相互作用的见解。