Ultraviolet photodissociation mass spectrometry of bis-aryl hydrazone conjugated peptides

Anal Chem. 2009 Jun 15;81(12):4864-72. doi: 10.1021/ac9005233.

Abstract

Ultraviolet photodissociation (UVPD) at 355 nm was used to rapidly identify peptides which had been chemically conjugated through bis-aryl hydrazone (BAH) moieties. The two biomolecules of interest were separately tagged to introduce either an aldehyde or a hydrazine and then conjugated together through these functional groups to from the UV-chromogenic BAH-group. In a mock mixture of peptides, UVPD was used to screen for the BAH-conjugated peptides in direct infusion ESI-UVPD-MS and online LC-UVPD-MS methods by comparing the abundances of the ions with the laser off and with the laser on. Only the BAH-conjugated peptides were observed to photodissociate upon exposure to UV irradiation, thus affording excellent selectivity for the pinpointing the relevant conjugated peptides in a complex mixture of nonconjugated peptides. UVPD analysis of conjugated model peptides indicated that the UVPD efficiencies of these species were charge state dependent. BAH-conjugated peptides that had a mobile proton which could protonate the basic BAH-moiety underwent more efficient photodissociation than the peptide ions with sequestered protons. Ultraviolet photodissociation of BAH-cross-linked peptides also yielded more diagnostic sequence ions than CID to unambiguously locate the site of conjugation.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Chromatography, Liquid
  • Hydrazones / chemical synthesis
  • Hydrazones / chemistry*
  • Mass Spectrometry / methods*
  • Peptides / analysis*
  • Peptides / chemistry
  • Photolysis
  • Spectrophotometry, Ultraviolet / methods*
  • Ultraviolet Rays*

Substances

  • Hydrazones
  • Peptides