The fate of b-ions in the two worlds of collision-induced dissociation

Biochim Biophys Acta. 2013 Dec;1834(12):2843-8. doi: 10.1016/j.bbapap.2013.08.007. Epub 2013 Aug 28.

Abstract

Fragment analysis of proteins and peptides by mass spectrometry using collision-induced dissociation (CID) revealed that the pairwise generated N-terminal b- and C-terminal y-ions have different stabilities resulting in underrepresentation of b-ions. Detailed analyses of large-scale spectra databases and synthetic peptides underlined these observations and additionally showed that the fragmentation pattern depends on utilized CID regime. To investigate this underrepresentation further we systematically compared resonant excitation energy and beam-type CID facilitated on different mass spectrometer platforms: (i) quadrupole time-of-flight, (ii) linear ion trap and (iii) three-dimensional ion trap. Detailed analysis of MS/MS data from a standard tryptic protein digest revealed that b-ions are significantly underrepresented on all investigated mass spectrometers. By N-terminal acetylation of tryptic peptides we show for the first time that b-ion cyclization reaction significantly contributes to b-ion underrepresentation even on ion trap instruments and accounts for at most 16% of b-ion loss.

Keywords: Ac; Collision-induced dissociation; ETD; HCD; HSD; LIT; Mass spectrometry; Peptide acetylation; Peptide fragmentation; QIT; QQQ; QToF; UPLC; acetylated; b-Ion cyclization; electron transfer dissociation; higher-energy collisional dissociation; honestly significant difference; linear ion trap; nAc; non-acetylated; quadrupole ion trap; quadrupole time-of-flight; triple quadrupole; ultra performance liquid chromatography.

MeSH terms

  • Animals
  • Aspergillus niger / enzymology
  • Bacterial Proteins / chemistry
  • Burkholderia / enzymology
  • Cattle
  • Databases, Protein*
  • Fungal Proteins / chemistry
  • Glucose Oxidase / chemistry
  • Lipase / chemistry
  • Mass Spectrometry / methods*
  • Peptides / chemistry*
  • alpha-Crystallins / chemistry

Substances

  • Bacterial Proteins
  • Fungal Proteins
  • Peptides
  • alpha-Crystallins
  • Glucose Oxidase
  • Lipase