Instrumentation for Analysis of High Mass Biomolecules

高质量生物分子分析仪器

• 批准号: 7021358
• 负责人: SCOTT A MCLUCKEY
• 金额: $ 24.81万
• 依托单位: PURDUE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 1991
• 资助国家: 美国
• 起止时间: 1991-09-01 至 2009-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7021358
• 关键词: biomedical equipment development; chemical association; electrospray ionization mass spectrometry; matrix assisted laser desorption ionization; oligonucleotides; protein structure function; structural biology

The objective of this work is to advance the state-of-the-art in whole protein mixture analysis with heavy emphasis on electrodynamic ion traps as reaction vessels for unimolecular and ion/ion reaction chemistries. Whole protein tandem mass spectrometry has been largely restricted to only a few types of mass spectrometry technologies and has, for the most part, been based on electrospray ionization. This proposal will emphasize the use of linear ion traps, which can serve as stand-alone mass analyzers or can readily be interfaced with other forms of mass analysis. Furthermore, one aspect of the proposal is devoted to the gasphase conversion of singly charged ions to multiply charged ions, which will facilitate whole protein tandem mass spectrometry using desorption ionization methods, which tend to form singly charged ions. Such a development would enable technologies that are well-suited to the spatial interrogation of surfaces to enjoy the structural tools now largely restricted to multiply charged ions derived via electrospray ionization. Other aspects of the proposal promise to expand the use of ion/ion reactions for mixture analysis and for the derivation of structrual information from whole protein ions, including post-translationally modified species. Furthermore, this work promises to make much more widely accessible means for studying whole proteins and protein complexes without prior recourse to enzymatic digestion. Particular emphasis will be placed on improved understanding and performance of electrospray ionization and matrix assisted laser desorption ionization applied to protein mixtures, charge state manipulation reactions, including charge inversion and, in some cases, sequential charge inversion steps, ion/ion reactions that provide structural information that is complementary to collisional activation of protonated species, and instrumentation development focused on stand-alone linear ion trap instrumentation and linear ion traps coupled with orthogonal acceleration time-of-flight mass analysis.

这项工作的目的是推进国家的最先进的全蛋白质混合物分析，重点是电动离子阱作为反应容器的单分子和离子/离子反应化学。全蛋白质串联质谱在很大程度上仅限于少数几种类型的质谱技术，并且在大多数情况下，基于电喷雾电离。该提案将强调使用线性离子阱，它可以作为独立的质量分析仪，也可以很容易地与其他形式的质量分析接口。此外，该提案的一个方面致力于单电荷离子到多电荷离子的气相转化，这将有助于使用解吸电离方法的全蛋白质串联质谱，该方法倾向于形成单电荷离子。这样的发展将使非常适合于表面的空间询问的技术能够享受现在主要限于通过电喷雾电离获得的多电荷离子的结构工具。该提案的其他方面有望扩大离子/离子反应用于混合物分析和从整个蛋白质离子（包括后修饰物种）中推导结构信息的用途。 此外，这项工作有望使更广泛的研究整个蛋白质和蛋白质复合物的手段，而无需事先求助于酶消化。 特别强调的是提高对应用于蛋白质混合物的电喷雾电离和基质辅助激光解吸电离的理解和性能、电荷状态操纵反应（包括电荷反转和在某些情况下连续的电荷反转步骤）、提供结构信息的离子/离子反应与质子化物质的碰撞激活互补，和仪器开发集中在独立的线性离子阱仪器和与正交加速飞行时间质量分析耦合的线性离子阱。