Search for the Structural Basis of Biomacromolecular Fun

寻找生物大分子乐趣的结构基础

• 批准号: 7338503
• 负责人: YUN WANG
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7338503
• 关键词: Search; Structural; Basis; Biomacromolecular; Fun

(1). We have completed the functional dynamics study of the L11-RNA-thiostrepton system, but decided not to pursue the structural determination of the ternary complex of L11-rRNA-thiostrepton as I have originally planed, because the result of our dynamics study appears to suggest that the structures of the ternary and the binary (L11-rRNA) complexes are similar. The results of the functional dynamics study has been written in a manuscript that was submitted to Journal of Molecular Biology. (2). We have nearly completed the "3P" (periodicity, Planarity and pixels) method development, and are in the process to program it in C++ in order to interface with the world-wide popular XPLOR-NIH package. The "3P" method was born out of a need to find a better way to determine structures like RAP that consists of periodical structural elements, and represents a significant departure from the traditional way of determining the structure using NMR data. One of the "3P" articles was selected for the cover page of Journal of Magnetic Resonance. A manuscript, the final installment of the "3P" project, has been written for a submission after the completion of the re-coding into C+++. This methodology development has facilitated our structural biology study of RAP, and will be useful for projects of structure determination in the future.The structural biology study of RAP and its interaction with the LRP receptor is an example of my vision as how a structural biology study should be conducted. This study tackled a difficult structural problem with a persistent effort, was aimed at the answering a fundamental biological question and was carried out with multiple collaborations. This project led to the discovery of a novel histidine switch that mediates the release of the LRP receptor in acidic environment in the Golgi, an important process involved in the receptor maturation. The mutants that we have made during the course of the study have potential therapeutic applications in treatment of diseases involved with the LRP receptor. We will apply for a US patent in the near future. The discovery of the histidine switch in RAP, a chaperone protein, trail blazes a new way to sequester the LRP receptor, and potentially other receptors, including LRP5 and LRP6 of the same family. Correct folding of LRP5/6 requires yet another specialized chaperone MESD. MESD can also be engineered to sequester LRP5/6 in the way similar to the mutated RAP to LRP, and this hypothesis sets a starting point to study the structural biology of the Wnt/beta-catenin signaling pathway. Looking into the future, to fulfill the research goal, I have setup several key collaborations with intramural and extramural groups. Among those, we plan to crystallize receptors or receptor fragments, whenever approiate, in collaboration with Dr. Ji's group; we plan to work with Dr. Shaprio's group to develop a new method for structure determination of large protein receptors and RNAmolecules; we plan to study the kinetic and thermodynamics in vitro of the interaction between the chaperones and the receptors in collaboration with Dr. Smith-Gill's group. We will continue our on-going collaboration with Professor Dudley Strickland group in the receptor biochemistry and biology.

(1).我们已经完成了L11-RNA-硫链丝菌肽系统的功能动力学研究，但决定不按照我最初的计划进行L11-rRNA-硫链丝菌肽三元复合物的结构测定，因为我们的动力学研究结果似乎表明三元和二元 (L11-rRNA) 复合物的结构相似。功能动力学研究的结果已写在提交给《分子生物学杂志》的手稿中。 （2）。我们已经接近完成了“3P”（周期性、平面性和像素）方法的开发，并且正在用 C++ 对其进行编程，以便与世界范围内流行的 XPLOR-NIH 软件包进行接口。 “3P”方法的诞生是为了找到一种更好的方法来确定结构，如由周期性结构元素组成的 RAP，它与使用 NMR 数据确定结构的传统方法有很大不同。其中一篇“3P”文章被选为《Journal of Magnetic Resonance》封面。手稿是“3P”项目的最后一部分，在完成重新编码为 C++ 后已完成提交。这种方法的发展促进了我们对 RAP 的结构生物学研究，并将对未来的结构测定项目有用。RAP 的结构生物学研究及其与 LRP 受体的相互作用是我对结构生物学研究的愿景的一个例子应进行。这项研究通过不懈的努力解决了一个困难的结构性问题，旨在回答一个基本的生物学问题，并通过多个合作进行。该项目发现了一种新型组氨酸开关，可介导高尔基体酸性环境中 LRP 受体的释放，这是受体成熟的一个重要过程。我们在研究过程中制造的突变体在治疗与 LRP 受体有关的疾病方面具有潜在的治疗应用。我们将在不久的将来申请美国专利。 RAP（一种伴侣蛋白）中组氨酸开关的发现开辟了一种隔离 LRP 受体以及潜在其他受体（包括同一家族的 LRP5 和 LRP6）的新方法。 LRP5/6 的正确折叠需要另一个专门的伴侣 MESD。 MESD 也可以通过与 RAP 突变为 LRP 类似的方式来隔离 LRP5/6，这一假设为研究 Wnt/β-catenin 信号通路的结构生物学奠定了起点。展望未来，为了实现研究目标，我与校内和校外团体建立了多项重要合作。其中，我们计划与季博士团队合作，在适当的时候结晶受体或受体片段；我们计划与Shaprio博士的团队合作，开发一种确定大蛋白受体和RNA分子结构的新方法；我们计划与 Smith-Gill 博士的团队合作，在体外研究伴侣与受体之间相互作用的动力学和热力学。我们将继续与 Dudley Strickland 教授小组在受体生物化学和生物学方面的持续合作。