Elucidating Factors Affecting the Preservation of Biomolecule Solution Structure for Spray-based Ionization Processes

阐明影响喷雾电离过程生物分子溶液结构保存的因素

• 批准号: 2305137
• 负责人: Stephen Valentine
• 金额: $ 39万
• 依托单位: West Virginia University Research Corporation
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-08-01 至 2026-07-31
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2305137&HistoricalAwards=false
• 关键词: Elucidating; Factors; Affecting; Preservation; Biomolecule

With support from the Chemical Measurement and Imaging Program in the Division of Chemistry, and the Established Program to Stimulate Competitive Research (EPSCoR), Stephen Valentine, Peng Li and Matthew Johnson and their groups at West Virginia University are developing new methods to study the structures of biological macromolecules such as proteins. These molecules must foldinto very specific three-dimensional (3D) structures in order to perform their required biological functions. Drs. Valentine, Li, and Johnson are developing new tools for generating micrometer-sized droplets of varying charge that allow the study of challenging biomolecules in solution using mass spectrometry (MS) techniques. The proposed studies provide fundamental knowledge required to conduct the most efficient MS measurements for biomolecule structure determination. In the future, this knowledge can be used to study disease processes as well as to develop biomolecular drugs. The work also provides valuable technology exposure to diverse graduate and undergraduate students in the areas of MS measurement and instrumentation development.The Valentine/Li/Johnson team at West Virginia University is developing a new approach to examine biomolecular structure using native MS. Recent experiments have shown that the technology of vibrating sharp-edge spray ionization (VSSI), when operated in a voltage-free mode for native MS experiments, can better preserve the structures of challenging proteins such as those possessing regions of disorder. For ion production, VSSI provides the ability to decouple the processes of droplet charging and droplet production in contrast to the Coloumbically-driven process encountered in electrospray ionization (ESI). Thus, with VSSI, it appears to now be possible to map protein structure transformations associated with specific droplet conditions. In this study the relationships between such transformations and droplet characteristics such as overall charge, diameter, and temperature equilibration with the heated capillary will be examined. The scientific knowledge gained from these studies will be shared publicly and can then be used to guide scientists with regard to best nanoelectropsray ionization (nESI) practices for studying the structures of challenging biomolecular species. In this way, the work has the potential for broad scientific impact across diverse fields from protein biophysics, to protein biocatalysis, to proteomics to human biology and biomedical science.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以及刺激竞争性研究的既定项目（EPSCoR）的支持下，西弗吉尼亚大学的Stephen Valentine、Peng Li和Matthew约翰逊及其团队正在开发研究蛋白质等生物大分子结构的新方法。 这些分子必须折叠成非常特定的三维（3D）结构，以执行其所需的生物功能。Valentine、Li和约翰逊博士正在开发新的工具，用于产生不同电荷的微米大小的液滴，从而可以使用质谱（MS）技术研究溶液中具有挑战性的生物分子。拟议的研究提供了进行生物分子结构测定的最有效的MS测量所需的基础知识。 在未来，这些知识可用于研究疾病过程以及开发生物分子药物。这项工作还为MS测量和仪器开发领域的不同研究生和本科生提供了有价值的技术接触。西弗吉尼亚大学的Valentine/Li/约翰逊团队正在开发一种新的方法来使用原生MS检查生物分子结构。最近的实验表明，振动锐边喷雾电离（VSSI）技术，当在原生MS实验的无电压模式下操作时，可以更好地保留具有挑战性的蛋白质的结构，例如那些具有无序区域的蛋白质。 对于离子产生，与电喷雾电离（ESI）中遇到的Coloumbically-driven过程相比，VSSI提供了将液滴充电和液滴产生的过程解耦的能力。 因此，使用VSSI，现在似乎可以映射与特定液滴条件相关的蛋白质结构转化。 在这项研究中，这种转换和液滴特性，如总电荷，直径和温度平衡与加热毛细管之间的关系将被检查。 从这些研究中获得的科学知识将公开分享，然后可以用来指导科学家关于最佳nanoelectropsray电离（nESI）的做法，研究具有挑战性的生物分子物种的结构。 通过这种方式，这项工作有可能在蛋白质生物物理学、蛋白质生物催化、蛋白质组学、人类生物学和生物医学等不同领域产生广泛的科学影响。该奖项反映了NSF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评审标准进行评估，被认为值得支持。