Next-Generation Collision Induced Unfolding Analysis of Proteins

下一代蛋白质碰撞诱导解折叠分析

• 批准号: 1808541
• 负责人: Brandon Ruotolo
• 金额: $ 33万
• 依托单位: Regents of the University of Michigan - Ann Arbor
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-08-01 至 2024-07-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=1808541&HistoricalAwards=false
• 关键词: Next; Generation; Collision; Induced; Unfolding

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, and with partial co-funding from the Division of Molecular and Cellular Biosciences, Professor Brandon Ruotolo and his group at the University of Michigan are developing powerful new tools for characterizing the 3-dimensional structure of proteins - key molecules in life processes. Specifically, the Ruotolo group is expanding the capabilities of "collision induced unfolding" (CIU), an approach which studies protein structures by quickly removing the water which surrounds protein molecules in their normal environment, then induces their unfolding while monitoring their size and shape using a state-of-the-art tool known as ion mobility-mass spectrometry (IM-MS). The approach has the potential to revolutionize the detail and acquisition speed with which protein stability can be evaluated. Potential applications include the rapid analysis of biotherapeutic proteins and their mode of operation. The work provides excellent training for students interested in research at the interface between chemical measurements and the pharmaceutical sciences.The research addresses three main goals: (1) creating a complete physical description of factors impacting the peak width for traveling-wave-type ion mobility separators, and validating the resulting predictions against standards; (2) developing next-generation software that incorporates the interpretation of IM peak width into CIU analysis workflows in the form of highly-parameterized multi-modal Gaussian fitting in both two and three dimensions; and (3) creating parallelized, multi-dimensional CIU methods capable of rapidly tracking the unfolding patterns of protein mixtures across many charge states simultaneously, even in the presence of significant chemical noise. The new CIU tools are being tested using therapeutic proteins of significance to the biotechnology sector.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.

在化学系化学测量和成像项目的支持下，以及分子和细胞生物科学系的部分共同资助下，密歇根大学的 Brandon Ruotolo 教授和他的团队正在开发强大的新工具来表征 3 - 蛋白质的维度结构 - 生命过程中的关键分子。 具体来说，Ruotolo 小组正在扩展“碰撞诱导展开”(CIU) 的能力，这种方法通过快速去除正常环境中蛋白质分子周围的水来研究蛋白质结构，然后诱导其展开，同时监测其大小和形状一种称为离子淌度质谱 (IM-MS) 的最先进工具。 该方法有可能彻底改变评估蛋白质稳定性的细节和采集速度。 潜在的应用包括生物治疗蛋白质及其操作模式的快速分析。 这项工作为对化学测量和制药科学之间的交叉研究感兴趣的学生提供了良好的培训。该研究解决了三个主要目标：(1) 对影响行波型离子淌度峰宽的因素创建完整的物理描述分隔符，并根据标准验证结果预测； (2) 开发下一代软件，以二维和三维高度参数化多模态高斯拟合的形式将 IM 峰宽的解释纳入 CIU 分析工作流程中； (3) 创建并行、多维 CIU 方法，即使在存在显着化学噪声的情况下，也能够同时快速跟踪多个电荷状态下蛋白质混合物的展开模式。 新的 CIU 工具正在使用对生物技术领域具有重要意义的治疗性蛋白质进行测试。该奖项反映了 NSF 的法定使命，并通过使用基金会的智力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

A Semi-Empirical Framework for Interpreting Traveling Wave Ion Mobility Arrival Time Distributions

解释行波离子淌度到达时间分布的半经验框架

• DOI: 10.1007/s13361-019-02133-6
• 发表时间: 2019-02-27
• 期刊: Journal of The American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: S. Dixit;B. Ruotolo
• 通讯作者: B. Ruotolo

Site-specific chirality-conferred structural compaction differentially mediates the cytotoxicity of Aβ42

位点特异性手性赋予的结构压实差异介导 Aβ42 的细胞毒性

• DOI: 10.1039/d3sc00678f
• 发表时间: 2023-06
• 期刊: Chemical Science
• 影响因子: 8.4
• 作者: Li, Gongyu;Jeon, Chae Kyung;Ma, Min;Jia, Yifei;Zheng, Zhen;Delafield, Daniel G.;Lu, Gaoyuan;Romanova, Elena V.;Sweedler, Jonathan V.;Ruotolo, Brandon T.;et al
• 通讯作者: et al

Comparing Selected-Ion Collision Induced Unfolding with All Ion Unfolding Methods for Comprehensive Protein Conformational Characterization

比较选择离子碰撞诱导解折叠与所有离子解折叠方法以进行全面的蛋白质构象表征

• DOI: 10.1021/jasms.2c00004
• 发表时间: 2022-06
• 期刊: Journal of the American Society for Mass Spectrometry
• 影响因子: 3.2
• 作者: Phetsanthad, Ashley;Li, Gongyu;Jeon, Chae Kyung;Ruotolo, Brandon T.;Li, Lingjun
• 通讯作者: Li, Lingjun

An Algorithm for Building Multi-State Classifiers Based on Collision-Induced Unfolding Data

一种基于碰撞诱导展开数据构建多状态分类器的算法

• DOI: 10.1021/acs.analchem.9b02650
• 发表时间: 2019-07
• 期刊: Analytical Chemistry
• 影响因子: 7.4
• 作者: Polasky, Daniel A.;Dixit, Sugyan M.;Vallejo, Daniel D.;Kulju, Kathryn D.;Ruotolo, Brandon T.
• 通讯作者: Ruotolo, Brandon T.

Bicarbonate buffers can promote crosslinking and alternative gas-phase dissociation pathways for multiprotein complexes

碳酸氢盐缓冲液可以促进多蛋白复合物的交联和替代气相解离途径

• DOI: 10.1016/j.ijms.2021.116687
• 发表时间: 2021-11
• 期刊: International Journal of Mass Spectrometry
• 影响因子: 1.8
• 作者: Li, Gongyu;Huang, Junfeng;Zheng, Zhen;Cao, Qinjingwen;Tian, Yuwei;Huang, Guangming;Li, Lingjun;Ruotolo, Brandon T.
• 通讯作者: Ruotolo, Brandon T.