Charge Site Mass Spectra: Conformation-Sensitive Components of the Electron Capture Dissociation Spectrum of a Protein

  • Owen S. Skinner
  • Kathrin Breuker
  • Fred W. McLafferty
Short Communication

Abstract

A conventional electron capture dissociation (ECD) spectrum of a protein is uniquely characteristic of the first dimension of its linear structure. This sequence information is indicated by summing the primary cm+ and zm+• products of cleavage at each of its molecular ion’s inter-residue bonds. For example, the ECD spectra of ubiquitin (M + nH)n+ ions, n = 7–13, provide sequence characterization of 72 of its 75 cleavage sites from 1843 ions in seven c(1–7)+ and eight z(1–8)+• spectra and their respective complements. Now we find that each of these c/z spectra is itself composed of “charge site (CS)” spectra, the cm+ or zm+• products of electron capture at a specific protonated basic residue. This charge site has been H-bonded to multiple other residues, producing multiple precursor ion forms; ECD at these residues yields the multiple products of that CS spectrum. Closely similar CS spectra are often formed from a range of charge states of ubiquitin and KIX ions; this indicates a common secondary conformation, but not the conventional α-helicity postulated previously. CS spectra should provide new capabilities for comparing regional conformations of gaseous protein ions and delineating ECD fragmentation pathways.

Key words

Electron capture dissociation (ECD) Charge site (CS) mass spectra Protein ion conformation 

Supplementary material

13361_2013_603_MOESM1_ESM.docx (3.4 mb)
ESM 1(DOCX 3479 kb)

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Copyright information

© American Society for Mass Spectrometry 2013

Authors and Affiliations

  • Owen S. Skinner
    • 1
  • Kathrin Breuker
    • 2
  • Fred W. McLafferty
    • 1
  1. 1.Department of Chemistry and Chemical BiologyCornell UniversityIthacaUSA
  2. 2.Institute of Organic Chemistry and Center for Molecular Biosciences InnsbruckUniversity of InnsbruckInnsbruckAustria

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