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

在化学系化学测量和成像计划的支持下，西弗吉尼亚大学的斯蒂芬·瓦伦丁、彭骊和马修·约翰逊及其团队正在开发新的方法来研究蛋白质等生物大分子结构。这些分子必须折叠成非常具体的三维(3D)结构，才能发挥其所需的生物学功能。瓦伦丁博士、李博士和约翰逊博士正在开发新的工具，用于产生微米大小的不同电荷的液滴，使使用质谱仪(MS)技术研究溶液中具有挑战性的生物分子成为可能。建议的研究提供了进行最有效的MS测量以确定生物分子结构所需的基础知识。在未来，这些知识可以用于研究疾病过程以及开发生物分子药物。这项工作还为不同的研究生和本科生在MS测量和仪器开发领域提供了宝贵的技术接触。西弗吉尼亚大学的瓦伦丁/李/约翰逊团队正在开发一种使用天然MS检查生物分子结构的新方法。最近的实验表明，振动尖端喷雾电离(VSSI)技术在天然MS实验的无电压模式下运行时，可以更好地保存具有挑战性的蛋白质的结构，例如那些具有无序区的蛋白质。对于离子生产，VSSI提供了将液滴充电和液滴生产过程分离的能力，而不是在电喷雾电离(ESI)中遇到的彩色驱动过程。因此，有了VSSI，现在似乎有可能绘制与特定液滴条件相关的蛋白质结构转换图。在这项研究中，将研究这种转变与液滴特性之间的关系，如总电荷、直径和与加热毛细管的温度平衡。从这些研究中获得的科学知识将被公开分享，然后可以用来指导科学家研究具有挑战性的生物分子结构的最佳纳米电离(NESI)实践。通过这种方式，这项工作有可能在从蛋白质生物物理学到蛋白质生物催化，再到蛋白质组学到人类生物学和生物医学科学的不同领域产生广泛的科学影响。这一奖项反映了NSF的法定使命，并通过使用基金会的智力优势和更广泛的影响审查标准进行评估，被认为值得支持。

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.