Type of Document Dissertation Author McFarland, Melinda A URN etd-08252004-191254 Title Fourier Transform Ion Cyclotron Resonance Mass Spectrometry Instrumentation and Methods for Structural Characterization of Trapped Biomolecular Ions: Innovative MS/MS Techniques, Gas-Phase Hudrogen/Deuterium Exchange, and Laser-Induced Fluoresnence Degree Doctor of Philosophy Department Chemistry and Biochemistry, Department of Advisory Committee
Advisor Name Title Alan G. Marshall Committee Chair Christopher L. Hendrickson Committee Co-Chair Naresh Dalal Committee Member Oliver Steinbock Committee Member Piotr G. Fajer Committee Member Keywords
- FT-ICR MS
- ion threshing
- gas phase HDX
Date of Defense 2004-08-16 Availability unrestricted AbstractFourier 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 is the optimization of the diverse range of fragmentation techniques available to FT-ICR MS/MS, including ion threshing, a novel form of collisionally activated dissociation (CAD), infrared multiphoton dissociation (IRMPD), electron capture dissociation (ECD), and electron detachment dissociation (EDD). Each of the dissociation techniques can be used separately, successively, or, in some cases, simultaneously to elucidate biomolecular sequences and modifications from their complementary fragmentation patterns. Millisecond timescale dissociation and mass selective ion accumulation facilitates MS/MS on a timescale compatible with continuous flow ionization methods. Also important are the powerful tools available with FT-ICR for the manipulation of trapped gas-phase ions.
Described here is the implementation, development and characterization of FT-ICR based MS/MS techniques for protein and peptide ion dissociation. 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. 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, as well as the novel application of ECD, EDD, and IRMPD to sulfatides and gangliosides. Also described is the use of gas-phase H/D exchange and fluorescence spectroscopy to probe conformation in gas-phase biological molecules.
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