CAREER: Dynamic Regulation of Protein Quality Control

职业：蛋白质质量控​​制的动态调节

• 批准号: 1552113
• 负责人: Richard Page
• 金额: $ 92.39万
• 依托单位: Miami University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-03-01 至 2019-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1552113&HistoricalAwards=false
• 关键词: CAREER; Dynamic; Regulation; Protein; Quality

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.

职务名称：职业：蛋白质质量控制的动态调节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。当暴露于化学或机械应力时，蛋白质可以错误折叠，导致三维结构的巨大变化和蛋白质功能的丧失。为了生存，细胞已经开发出质量控制系统，引导错误折叠的蛋白质进入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指导错误折叠的蛋白质进行破坏来响应蛋白质错误折叠的生物学原理。该项目的另一个目标是开发技术，以进一步研究如何控制细胞对蛋白质错误折叠的反应。这个职业项目的综合教育目标是通过直接推广高中和本科阶段，通过实践研究经验和不同的推广计划，增加在STEM中代表性不足的少数民族的保留。该项目将直接将研究融入研究生和本科生课程，重点是对真实的数据的实际解释，并关注视觉，口头和书面沟通技能的发展。细胞对机械、热或化学应力的反应在生物学中是普遍存在的。E3泛素连接酶CHIP和70千道尔顿的热休克蛋白Hsp 70通过融合重折叠和降解途径在细胞应激反应中发挥关键作用。这项研究将建立皮秒到毫秒时间尺度上的动力学与陪伴泛素化复合物的生物功能之间的联系。利用高分辨率X射线晶体学信息开发的功能获得性突变体的掺入将确定增强CHIP/Hsp 70相互作用的效果。该项目将重新定义蛋白质质量控制领域如何看待Hsp 70和CHIP之间的相互作用，通过采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。NMR、SAXS和EPR与生物层干涉测量的结合将在结构生物学的混合方法的使用中产生进步。一种新的实时泛素化检测将证明生物层干涉法的实用性，快速和灵敏的监测进行性翻译后修饰。

项目成果

Using PyMOL to Explore the Effects of pH on Noncovalent Interactions between Immunoglobulin G and Protein A: A Guided-Inquiry Biochemistry Activity: Guided Inquiry Activity: PyMOL to Explore Protein Interactions

使用 PyMOL 探索 pH 对免疫球蛋白 G 和蛋白 A 之间非共价相互作用的影响：引导探究生化活动：引导探究活动：PyMOL 探索蛋白质相互作用

• DOI: 10.1002/bmb.21066
• 发表时间: 2017
• 期刊: Biochemistry and Molecular Biology Education
• 影响因子: 1.4
• 作者: Roche Allred, Zahilyn D.;Tai, Heeyoung;Bretz, Stacey Lowery;Page, Richard C.
• 通讯作者: Page, Richard C.