Characteristic fragment ions associated with dansyl cadaverine and biotin cadaverine adducts on glutamine

Anal Biochem. 2020 Jul 1:600:113718. doi: 10.1016/j.ab.2020.113718. Epub 2020 Apr 23.

Abstract

Glutamine residues susceptible to transglutaminase-catalyzed crosslinking can be identified by incorporation of dansyl cadaverine or biotin cadaverine. Bacterial transglutaminase and human transglutaminase 2 were used to modify residues in beta-casein with dansyl cadaverine. Bacterial transglutaminase was used to modify residues in human butyrylcholinesterase with biotin cadaverine. Tryptic peptides were analyzed by LC-MS/MS on an Orbitrap Fusion Lumos mass spectrometer. Modified residues were identified in Protein Prospector searches of mass spectrometry data. The MS/MS spectra from modified casein included intense peaks at 336.2, 402.2, and 447.2 for fragments of dansyl cadaverine adducts on glutamine. The MS/MS spectra from modified butyrylcholinesterase included intense peaks at 329.2, 395.2, and 440.2 for fragments of biotin cadaverine adducts on glutamine. No evidence for transglutaminase-catalyzed adducts on glutamic acid, aspartic acid, or asparagine was found. Consistent with expectation, it was concluded that bacterial transglutaminase and human transglutaminase 2 specifically modify glutamine. The characteristic ions associated with dansyl cadaverine and biotin cadaverine adducts on glutamine are useful markers for modified peptides.

Keywords: Biotin cadaverine; Butyrylcholinesterase; Casein; Dansyl cadaverine; Mass spectrometry; Protein prospector; Transglutaminase.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Biotin / chemistry*
  • Biotin / metabolism
  • Butyrylcholinesterase / metabolism
  • Cadaverine / chemistry*
  • Cadaverine / metabolism
  • Glutamine / chemistry*
  • Glutamine / metabolism
  • Humans
  • Ions / chemistry
  • Ions / metabolism
  • Streptomyces / enzymology
  • Transglutaminases / metabolism

Substances

  • Ions
  • Glutamine
  • Biotin
  • Transglutaminases
  • Butyrylcholinesterase
  • Cadaverine

Supplementary concepts

  • Streptomyces mobaraensis