CAREER: Dynamic Regulation of Protein Quality Control

蛋白质的三维结构决定了蛋白质在细胞内发挥的作用和功能。在暴露于化学或机械应力下，蛋白质会发生错误折叠，导致三维结构的巨大变化和蛋白质功能的丧失。为了生存，细胞已经发展出质量控制系统，引导错误折叠的蛋白质走向导致它们被修复或丢弃的途径。该项目将确定允许细胞通过引导错误折叠的蛋白质进行破坏来对蛋白质错误折叠做出反应的生物学原理。该项目的另一个目标是开发能够进一步研究如何控制细胞对蛋白质错误折叠的反应的技术。这个CAREER项目的综合教育目标是通过在高中和本科阶段的直接推广，通过实践研究经验和不同的推广项目，增加STEM中代表性不足的少数民族的保留率。该项目将把研究直接整合到研究生和本科生课程中，重点是对真实数据的动手解释，并关注视觉、口头和书面沟通技巧的发展。细胞对机械、热或化学压力的反应需求在生物学中是普遍存在的。E3泛素连接酶CHIP和70千dalton热休克蛋白Hsp70通过重折叠和降解途径的收敛在细胞应激反应中发挥关键作用。该研究将建立皮秒到毫秒时间尺度上的动力学与伴随泛素化复合物的生物学功能之间的联系。利用高分辨率x射线晶体学信息开发的功能获得突变体的结合将确定增强CHIP/Hsp70相互作用的影响。该项目将重新定义蛋白质质量控制领域如何看待Hsp70和CHIP之间的相互作用，通过使用重折叠或降解方法来调节细胞对蛋白质错误折叠的反应。核磁共振、SAXS和EPR与生物层干涉测量的结合将在结构生物学中使用混合方法方面取得进展。一种新型的实时泛素化分析将展示生物层干涉测量法的实用性，用于快速和敏感地监测过程中的翻译后修饰。

Title: CAREER: Dynamic Regulation of Protein Quality ControlThe three-dimensional structures of proteins determine the roles and functions proteins play within cells. Upon exposure to chemical or mechanical stresses, proteins can misfold, resulting in large changes in three-dimensional structure and loss of protein function. To survive, cells have developed quality control systems that guide misfolded proteins towards pathways that lead to them either being repaired or discarded. This project will determine the biological principles that allow cells to respond to protein misfolding by directing misfolded proteins for destruction. Another goal of this project is to develop techniques that will enable further investigations into how cellular responses to protein misfolding can be controlled. The integrated education objective of this CAREER project is to increase retention of underrepresented minorities in STEM through direct outreach at the high school and undergraduate levels via hands-on research experience and through different outreach programs. The project will integrate research directly into graduate and undergraduate courses with an emphasis on hands-on interpretation of real data and attention to development of visual, verbal and written communication skills. The requirement for cells to respond to mechanical, thermal or chemical stresses is ubiquitous in biology. The E3 ubiquitin ligase CHIP and the 70 kilodalton heat shock protein, Hsp70, play pivotal roles in the response to cellular stress through the convergence of refolding and degradative pathways. The research will forge connections between dynamics on the picosecond to millisecond timescales and the biological function of chaperoned ubiquitination complexes. The incorporation of gain-of-function mutants developed using insights from high-resolution X-ray crystallographic information will identify the effects of enhancing the CHIP/Hsp70 interaction. The project will redefine how the protein quality control field views the role of interactions between Hsp70 and CHIP in regulating how cells respond to protein misfolding by employing either refolding or degradative approaches. The combination of NMR, SAXS, and EPR with biolayer interferometry will generate advances in the use of hybrid methods for structural biology. A novel real-time ubiquitination assay will demonstrate the utility of biolayer interferometry for rapid and sensitive monitoring of a processive post-translational modification.