<title>Correlation between solution and gas-phase protein conformation: H/D exchange, IRMPD, and ESI FT-ICR MS</title>
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Infrared multiphoton dissociation (IRMPD) of the hydrogen/deuterium (H/D) exchanged 12+ charge state of gas-phase bovine ubiquitin was performed on a Fourier transform ion cyclotron resonance mass spectrometer. The H/D exchange of the 12+ charge state revealed two distinct isotopic distributions indicating the presence of at least two distinct conformations of the 12+ charge state. Following H/D exchange, IRMPD was used to dissociate the conformations. The fragments clearly showed little or no deuterium scrambling as evidenced by a nonstatistical distribution of deuterium incorporation. Analysis of the deuterium incorporation for the five most abundant fragment ions indicated a slow exchanging region of the fast exchanging conformation that corresponds to a stable (beta) - sheet observed by NMR in alcoholic solutions. The data suggest that protection of the amide hydrogens in the (beta) -sheet may result in the observed slow exchange rate and provides further evidence for the retention of secondary structure in gas phase.Keywords:
Infrared multiphoton dissociation
Hydrogen–deuterium exchange
Ion cyclotron resonance
An electron injection system based on an indirectly heated ring-shaped dispenser cathode has been developed and installed in a 7 Tesla Fourier transform ion cyclotron resonance (FTICR) mass spectrometer. This new hardware design allows high-rate electron capture dissociation (ECD) to be carried out by a hollow electron beam coaxial with the ion cyclotron resonance (ICR) trap. Infrared multiphoton dissociation (IRMPD) can also be performed with an on-axis IR-laser beam passing through a hole at the centre of the dispenser cathode. Electron and photon irradiation times of the order of 100 ms are required for efficient ECD and IRMPD, respectively. As ECD and IRMPD generate fragments of different types (mostly c, z and b, y, respectively), complementary structural information that improves the characterization of peptides and proteins by FTICR mass spectrometry can be obtained. The developed technique enables the consecutive or simultaneous use of the ECD and IRMPD methods within a single FTICR experimental sequence and on the same ensemble of trapped ions in multistage tandem (MS/MS/MS or MS(n)) mass spectrometry. Flexible changing between ECD and IRMPD should present advantages for the analysis of protein digests separated by liquid chromatography prior to FTICRMS. Furthermore, ion activation by either electron or laser irradiation prior to, as well as after, dissociation by IRMPD or ECD increases the efficiency of ion fragmentation, including the w-type fragment ion formation, and improves sequencing of peptides with multiple disulfide bridges. The developed instrumental configuration is essential for combined ECD and IRMPD on FTICR mass spectrometers with limited access into the ICR trap.
Infrared multiphoton dissociation
Electron-capture dissociation
Ion cyclotron resonance
Fourier transform spectroscopy
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Detachment of heme prosthetic groups from gaseous myoglobin ions has been studied by collision-induced dissociation and infrared multiphoton dissociation in combination with Fourier-transform ion cyclotron resonance mass spectrometry. Multiply charged holomyoglobin ions (hMbn+) were generated by electrospray ionization and transferred to an ion cyclotron resonance cell, where the ions of interest were isolated and fragmented by either collision with Ar atoms or irradiation with 3μm photons, producing apomyoglobin ions (aMbn+). Both charged heme loss (with [Fe(III)-heme]+ and aMb(n−1)+ as the products) and neutral heme loss (with [Fe(II)-heme] and aMbn+ as the products) were detected concurrently for hMbn+ produced from a myoglobin solution pretreated with reducing reagents. By reference to Ea=0.9eV determined by blackbody infrared radiative dissociation for charged heme loss of ferric hMbn+, an activation energy of 1.1eV was deduced for neutral heme loss of ferrous hMbn+ with n=9 and 10.
Infrared multiphoton dissociation
Ion cyclotron resonance
Fourier transform spectroscopy
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ADVERTISEMENT RETURN TO ISSUEPREVAdditions & Corr...Additions & CorrectionsORIGINAL ARTICLEThis notice is a correctionErratum to: An antibiotic linked to peptides and proteins is released by electron capture dissociation fourier transform ion cyclotron resonance mass spectrometryClifton K. FagerquistClifton K. FagerquistMore by Clifton K. Fagerquist, Robert R. HudginsRobert R. HudginsMore by Robert R. Hudgins, Mark R. EmmettMark R. EmmettMore by Mark R. Emmett, Kristina HåkanssonKristina HåkanssonMore by Kristina Håkansson, and Alan G. MarshallAlan G. MarshallMore by Alan G. MarshallCite this: J Am Soc Mass Spectrom 2003, 14, 6, 682Publication Date (Web):June 1, 2003Publication History Published online1 June 2003Published inissue 1 June 2003https://pubs.acs.org/doi/10.1016/S1044-0305(03)00331-3https://doi.org/10.1016/S1044-0305(03)00331-3correctionACS PublicationsCopyright © 2003 © American Society for Mass Spectrometry 2003. This publication is available under these Terms of Use. Request reuse permissions This publication is free to access through this site. Learn MoreArticle Views11Altmetric-Citations2LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InRedditEmail PDF (35 KB) Get e-Alerts
Electron-capture dissociation
Infrared multiphoton dissociation
Ion cyclotron resonance
Top-down proteomics
Selected ion monitoring
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Ion Structure and Energetics in the Gas Phase Characterized Using Fourier Transform Ion Cyclotron Resonance Mass Spectrometry Chad A. Jones Department of Chemistry and Biochemistry, BYU Doctor of Philosophy In this dissertation, I use Fourier transform ion cyclotron resonance mass spectrometry (FTICR-MS) to study the structure and energetics of gas phase ions. Infrared multiphoton dissociation spectroscopy (IRMPD) is a technique for measuring the IR spectrum of gas phase ions in a Penning trap. I use this technique to investigate the conformation of cucurbituril complexes, terminal diamines, and protonated amino acids. Cross sectional areas by Fourier transform ion cyclotron resonance mass spectrometry (CRAFTI) is a technique developed by the Dearden lab to measure the cross section of gas phase ions. In this work, I further develop a fundamental understanding of this technique. I investigate the role that dissociation plays in this and other FTICR-MS techniques. I also show that the principles of the CRAFTI technique can be used to measure the pressure inside the cell of an FTICR-MS. This technique, linewidth pressure measurement (LIPS), allows for a quantitative measurement and comparison of CRAFTI cross sections. To demonstrate the improvements to the technique, I measure the CRAFTI cross sections for the 20 standard amino acids and compare these to literature values measured by ion mobility measurements.
Infrared multiphoton dissociation
Ion cyclotron resonance
Selected ion monitoring
Top-down proteomics
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Electron-capture dissociation
Infrared multiphoton dissociation
Ion cyclotron resonance
Posttranslational modification
Top-down proteomics
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Abstract A novel set‐up for Fourier transform ion cyclotron resonance mass spectrometry (FTICR) is reported for simultaneous infrared multiphoton dissociation (IRMPD) and electron‐capture dissociation (ECD). An unmodified electron gun ensures complete, on‐axis overlap between the electron and the photon beams. The instrumentation, design and implementation of this novel approach are described. In this configuration the IR beam is directed into the ICR cell using a pneumatically actuated mirror inserted into the ion‐optical path. Concept validation was made using different combinations of IRMPD and ECD irradiation events on two standard peptides. The ability to perform efficient IRMPD, ECD and especially simultaneous IRMPD and ECD using lower irradiation times is demonstrated. The increase in primary sequence coverage, with the combined IRMPD and ECD set‐up, also increases the confidence in peptide and protein assignments. Copyright © 2006 John Wiley & Sons, Ltd.
Infrared multiphoton dissociation
Electron-capture dissociation
Ion cyclotron resonance
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Infrared multiphoton dissociation
Ion cyclotron resonance
Fourier transform spectroscopy
Cyclotron resonance
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Electron-capture dissociation
Ion cyclotron resonance
Infrared multiphoton dissociation
Top-down proteomics
Selected ion monitoring
Cyclotron resonance
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Infrared multiphoton dissociation (IRMPD) is a vibrational excitation tandem mass spectrometric fragmentation method valuable for sequencing of oligonucleotides. For oligodeoxynucleotides, typical product ions correspond to sequence-specific 5' (a-base) and their complementary 3' w-type ions from carbon-oxygen bond cleavage at the 3' position of the deoxyribose from which a nucleobase is lost. Such fragmentation patterns are also observed in collision activated dissociation (CAD). The CAD oligodeoxynucleotide fragmentation mechanism has been characterized in detail. By contrast, fragmentation schemes in IRMPD have not been rigorously established. In this paper, we apply, for the first time, Fourier transform ion cyclotron double resonance (DR) experiments to characterize IRMPD fragmentation pathways of oligodeoxynucleotide anions. Our results suggest that neutral base loss precedes backbone fragmentation but that T-rich oligodeoxynucleotides fragment via a different mechanism, similar to the mechanisms proposed for CAD. We also extend the DR approach to characterize intermediates in electron detachment dissociation of hexamer oligodeoxynucleotides. Here, we found that charge reduced radical precursor ions constitute major intermediates for dT(6), d(GCATAC) and d(GCATGC). Furthermore, (a/z-T) ions (z ions correspond to C-O bond cleavage on the other side of a backbone phosphate group as compared to the formation of a ions) mainly originate from secondary fragmentation of a/z radical ions for the oligodeoxynucleotide dT(6).
Infrared multiphoton dissociation
Fragmentation
Ion cyclotron resonance
Electron-capture dissociation
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