SITS Derivatization of Peptides to Enhance 266 nm Ultraviolet Photodissociation (UVPD)

  • M. Montana Quick
  • M. Rachel Mehaffey
  • Robert W. Johns
  • W. Ryan Parker
  • Jennifer S. Brodbelt
Focus: Bio-Ion Chemistry: Interactions of Biological Ions with Ions, Molecules, Surfaces, Electrons, and Light: Research Article

Abstract

N-terminal derivatization of peptides with the chromogenic reagent 4-acetamido-4-isothiocyanatostilbene-2,2-disulfonic acid (SITS) is demonstrated to enhance the efficiency of 266 nm ultraviolet photodissociation (UVPD). Attachment of the chromophore results in a mass shift of 454 Da and provides significant gains in the number and abundances of diagnostic fragment ions upon UVPD. Activation of SITS-tagged peptides with 266 nm UVPD leads to many fragment ions akin to the a/b/y ions commonly produced by CID, along with other sequence ions (c, x, and z) typically accessed through higher energy pathways. Extreme bias towards C-terminal fragment ions is observed upon activation of SITS-tagged peptides using multiple 266 nm laser pulses. Due to the high reaction efficiency of the isothiocyanate coupling to the N-terminus of peptides, we demonstrate the ability to adapt this strategy to a high-throughput LC-MS/MS workflow with 266 nm UVPD.

Graphical Abstract

Keywords

Ion activation Ultraviolet photodissociation Derivatization Peptide 

Notes

Acknowledgements

The authors acknowledge the following funding sources: NSF (Grant CHE1402753) and the Welch Foundation (Grant F-1155). R.W.J. acknowledges an ALS Doctoral Fellowship in Residence and support from the Welch Foundation (F-1848 to Delia Milliron).

Supplementary material

13361_2017_1650_MOESM1_ESM.pdf (541 kb)
ESM 1 (PDF 540 kb)

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Copyright information

© American Society for Mass Spectrometry 2017

Authors and Affiliations

  • M. Montana Quick
    • 1
  • M. Rachel Mehaffey
    • 1
  • Robert W. Johns
    • 2
    • 3
  • W. Ryan Parker
    • 1
  • Jennifer S. Brodbelt
    • 1
  1. 1.Department of ChemistryUniversity of TexasAustinUSA
  2. 2.Department of ChemistryUniversity of CaliforniaBerkeleyUSA
  3. 3.McKetta Department of Chemical EngineeringUniversity of TexasAustinUSA

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