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首页> 外文学位 >Fourier transform ion cyclotron resonance mass spectrometry instrumentation and methods for structural characterization of trapped biomolecular ions: Innovative MS/MS techniques, gas-phase hydrogen /deuterium exchange, and laser -induced fluorescence.
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Fourier transform ion cyclotron resonance mass spectrometry instrumentation and methods for structural characterization of trapped biomolecular ions: Innovative MS/MS techniques, gas-phase hydrogen /deuterium exchange, and laser -induced fluorescence.

机译：傅里叶变换离子回旋共振质谱仪和捕获的生物分子离子的结构表征方法：创新的MS / MS技术，气相氢/氘交换和激光诱导的荧光。

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摘要

Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR MS) has become a powerful tool for biomolecular structural characterization because of its superior mass resolving power and high mass accuracy. Of particular interest in this work are the diverse range of fragmentation techniques available to FT-ICR MS/MS, including collisionally activated dissociation (CAD), infrared multiphoton dissociation (IRMPD), electron capture dissociation (ECD), and electron detachment dissociation (EDD). Described here is the implementation, development and characterization of FT-ICR based MS/MS techniques for protein and peptide ion dissociation. Also described is the use of gas-phase H/D exchange and fluorescence spectroscopy to probe conformation in gas-phase biological molecules.;Implementation of efficient and rapid collisionally-activated dissociation (CAD) external to an ICR cell (ion threshing) by use of a novel axial electric potential gradient mounted in an external ion accumulation octopole is discussed. Rapid switching of the wire potential between a positive and negative value drives the ion axial motion back and forth and, in the presence of nitrogen gas at suitable pressure, induces dissociation. The method is sufficiently rapid for MS/MS with on-line liquid chromatography (LC) sample introduction. Moreover, compared to CAD in the ICR cell, external CAD improves mass accuracy, producing thermal on-axis fragment ions for detection.;Design and implementation of a permanent on-axis dispenser cathode electron source and off-axis laser geometry that enables simultaneous access to ECD and IRMPD in a 9.4 Tesla FT-ICR mass spectrometer is described. Optimum performance of both fragmentation techniques is maintained.;Infrared and electron irradiation dissociation techniques are primarily applied to structural elucidation of peptides and proteins. The novel application of ECD, EDD, and IRMPD to sulfatides and gangliosides is discussed. These techniques provide extensive structural information for both the ceramide and glycosyl regions of these glycosphingolipids. Also described is the use of mass defect graphical analysis for the simplification of identification of fragment ions. The substantial difference in mass defect between the hydrocarbon and saccharide regions graphically separate into classes.;The development of instrumentation and implementation of electron induced (EIF), and laser induced fluorescence (LIF) and lifetime measurements of mass selected trapped ions is also described. This work includes description of experimental apparatus, implementation of quadrupolar excitation and axialization (QE) for mass selection and ion axialization, as well as results for the EIF, LIF, and lifetime measurements for the trifluorobenzene cation. (Abstract shortened by UMI.).
机译：傅立叶变换离子回旋共振质谱（FT-ICR MS）由于其卓越的质量分辨能力和高质量精度，已成为生物分子结构表征的强大工具。这项工作中特别令人感兴趣的是FT-ICR MS / MS可用的各种裂解技术，包括碰撞活化解离（CAD），红外多光子解离（IRMPD），电子捕获解离（ECD）和电子离解解离（EDD） ）。本文描述了基于FT-ICR的MS / MS技术用于蛋白质和肽离子解离的实现，开发和表征。还描述了使用气相H / D交换和荧光光谱法探测气相生物分子中的构象。通过使用ICR细胞外部有效而快速的碰撞活化解离（CAD）（离子脱粒）讨论了安装在外部离子累积八极杆上的新型轴向电势梯度。导线电势在正值和负值之间快速切换会驱动离子轴向运动来回移动，并且在存在氮气的情况下，在合适的压力下会引起离解。对于采用在线液相色谱（LC）样品引入的MS / MS，该方法足够快速。此外，与ICR电池中的CAD相比，外部CAD改善了质量准确性，产生了热轴向碎片离子以进行检测;永久性同轴分配器阴极电子源和离轴激光几何结构的设计和实现，可同时访问描述了在9.4 Tesla FT-ICR质谱仪中的ECD和IRMPD。两种片段化技术均能保持最佳性能。红外和电子辐射解离技术主要用于肽和蛋白质的结构解析。讨论了ECD，EDD和IRMPD在硫化物和神经节苷脂上的新应用。这些技术为这些糖鞘脂的神经酰胺和糖基区域提供了广泛的结构信息。还描述了使用质量缺陷图形分析简化碎片离子的鉴定。烃区和糖区之间质量缺陷的实质差异以图形方式分为几类。还描述了电子诱导（EIF）和激光诱导荧光（LIF）的仪器和实现的发展以及对所选离子质量的寿命测量。这项工作包括实验设备的说明，用于质量选择和离子轴向化的四极激发和轴向化（QE）的实现，以及EIF，LIF的结果以及三氟苯阳离子的寿命测量。 （摘要由UMI缩短。）。

著录项

  • 作者

    McFarland, Melinda A.;

  • 作者单位

    The Florida State University.;

  • 授予单位 The Florida State University.;
  • 学科 Chemistry Physical.;Chemistry Analytical.;Chemistry Biochemistry.
  • 学位 Ph.D.
  • 年度 2004
  • 页码 188 p.
  • 总页数 188
  • 原文格式 PDF
  • 正文语种 eng
  • 中图分类
  • 关键词

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