How do proteins fold: Mechanism and principles

蛋白质如何折叠：机制和原理

• 批准号: 1929671
• 负责人: S. Englander
• 金额: $ 60万
• 依托单位: University of Pennsylvania
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-01 至 2023-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1929671&HistoricalAwards=false
• 关键词: How; do; proteins; fold; Mechanism

This project will investigate the mechanisms and principles that drive protein folding. Protein folding is fundamental to all living organisms. The pathway of intermediate structures that carry proteins from their initially synthesized unstructured condition to their biologically functional structure is arguably the most centrally important reaction in all of biology. Further, it is key to understanding many normal processes and to the effort to interdict challenging misfolding processes. The folding problem has proven extremely difficult. All available methodologies have been directed at the problem but they commonly cannot describe detailed structure formation during the kinetic folding process. This laboratory has developed and recently demonstrated a new approach, named hydrogen exchange analyzed by mass spectrometry (HX-MS), that shows great promise for delving into and perhaps solving this long-lasting enigma. The understanding of the folding process may provide the knowledge necessary to interfere in a constructive way with misfolding processes. The investigator has a superb record of training graduate students and postdocs and will continue these training activities. In addition, the proposed method development and software tools are likely to benefit the scientific community at large. Recent work in this laboratory has developed the pulse labeling HX MS method and shown it can be used to define, for the first time, the structure of the increasingly folded intermediate forms that together construct the protein folding pathway. Results so far obtained for three proteins (ribonuclease H, cytochrome c, and maltose binding protein) clearly display the timed development of structures that carry the initially unfolded proteins (U) through intermediate forms (Ii) to their final native state (N). Further the results suggest three principles that appear to govern the folding process. The plan going forward is to test for the reality and generality of these results. Initial efforts will be directed at the alpha-subunit of tryptophan synthase, a moderate-sized protein with 265 residues in a TIM barrel structure, which represents the most common protein fold, accounting for ~10% of all known proteins. The investigation will then be extended to the small all-beta protein ubiquitin (76 residues) and then ultimately to the immense Hsp104 protomer (908 residues).This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

这个项目将研究驱动蛋白质折叠的机制和原理。蛋白质折叠是所有生物体的基础。中间结构的途径将蛋白质从最初合成的非结构化状态携带到其生物功能结构，可以说是所有生物学中最重要的反应。此外，它是理解许多正常过程和努力阻止具有挑战性的错误折叠过程的关键。折叠问题已被证明是极其困难的。所有可用的方法都针对这个问题，但它们通常不能描述动力学折叠过程中的详细结构形成。该实验室已经开发并最近展示了一种名为质谱分析氢交换（HX-MS）的新方法，该方法有望深入研究并可能解决这个长期存在的谜题。对折叠过程的理解可以提供必要的知识，以建设性的方式干预错误的折叠过程。研究者在培养研究生和博士后方面有着出色的记录，并将继续这些培训活动。此外，所提出的方法开发和软件工具可能会使整个科学界受益。该实验室最近的工作开发了脉冲标记HX质谱方法，并首次表明它可以用来定义越来越多的折叠中间形式的结构，这些中间形式共同构建蛋白质折叠途径。迄今为止获得的三种蛋白质（核糖核酸酶H、细胞色素c和麦芽糖结合蛋白）的结果清楚地显示了结构的定时发展，这些结构将最初未折叠的蛋白质(U)通过中间形式（Ii）携带到最终的天然状态(N)。进一步的结果表明，三个原则似乎支配着折叠过程。接下来的计划是测试这些结果的真实性和普遍性。最初的努力将针对色氨酸合成酶的α亚基，这是一种中等大小的蛋白质，在TIM桶状结构中有265个残基，它代表了最常见的蛋白质折叠，占所有已知蛋白质的10%左右。然后，研究将扩展到小的全β蛋白泛素（76个残基），然后最终扩展到巨大的Hsp104原聚体（908个残基）。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

项目成果

ROR and RYK extracellular region structures suggest that receptor tyrosine kinases have distinct WNT-recognition modes.

• DOI: 10.1016/j.celrep.2021.109834
• 发表时间: 2021-10-19
• 期刊: Cell reports
• 影响因子: 8.8
• 作者: Shi F;Mendrola JM;Sheetz JB;Wu N;Sommer A;Speer KF;Noordermeer JN;Kan ZY;Perry K;Englander SW;Stayrook SE;Fradkin LG;Lemmon MA
• 通讯作者: Lemmon MA

ROR and RYK extracellular region structures suggest that receptor tyrosine kinases have distinct WNT-recognition modes

ROR 和 RYK 胞外区结构表明受体酪氨酸激酶具有不同的 WNT 识别模式

• DOI: 10.1101/2021.04.29.442059
• 发表时间: 2021
• 期刊: bioRxiv
• 作者: Fumin Shi1, 2;Lemmon1, 2
• 通讯作者: Lemmon1, 2

Structural and kinetic basis for the regulation and potentiation of Hsp104 function

Hsp104 功能调节和增强的结构和动力学基础

• DOI: 10.1073/pnas.192196811
• 发表时间: 2020
• 期刊: PNAS nexus
• 作者: Xiang Ye https:/;S. Walter Englander
• 通讯作者: S. Walter Englander

Hydrogen exchange reveals Hsp104 architecture, structural dynamics, and energetics in physiological solution

• DOI: 10.1073/pnas.1816184116
• 发表时间: 2019-03
• 期刊: Proceedings of the National Academy of Sciences
• 作者: X. Ye;JiaBei Lin;L. Mayne;J. Shorter;S. Englander