Characterization of Antigenic Oligosaccharides from Gram-Negative Bacteria via Activated Electron Photodetachment Mass Spectrometry

Anal Chem. 2019 Apr 2;91(7):4672-4679. doi: 10.1021/acs.analchem.9b00048. Epub 2019 Mar 15.

Abstract

Lipooligosaccharides (LOS), composed of hydrophilic oligosaccharides and hydrophobic lipid A domains, are found on the outer membranes of Gram-negative bacteria. Here we report the characterization of deacylated LOS of LPS by activated-electron photodetachment mass spectrometry. Collision induced dissociation (CID) of these phosphorylated oligosaccharides produces simple MS/MS spectra with most fragment ions arising from cleavages near the reducing end of the molecule where the phosphate groups are located. In contrast, 193 nm ultraviolet photodissociation (UVPD) generates a wide array of product ions throughout the oligosaccharide including cross-ring fragments that illuminate the branching patterns. However, there are also product ions that are redundant or uninformative, resulting in more congested spectra that complicate interpretation. In this work, a hybrid UVPD-CID approach known as activated-electron photodetachment (a-EPD) affords less congested spectra than UVPD alone and richer fragmentation patterns than CID alone. a-EPD combines UVPD of negatively charged oligosaccharides to yield abundant charge-reduced radical ions which are subsequently interrogated by collisional activation. CID of the charge-reduced precursors results in extensive fragmentation throughout the backbone of the oligosaccharide. This hybridized a-EPD approach was employed to characterize the structure and branching pattern of deacylated LOS of E. coli.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antigens / chemistry
  • Antigens / immunology
  • Electrons
  • Escherichia coli / immunology
  • Escherichia coli / metabolism*
  • Lipopolysaccharides / analysis*
  • Lipopolysaccharides / immunology
  • Lipopolysaccharides / metabolism
  • Spectrometry, Mass, Electrospray Ionization / methods*

Substances

  • Antigens
  • Lipopolysaccharides
  • lipid-linked oligosaccharides