Combining FAIMS based glycoproteomics and DIA proteomics reveals widespread proteome alterations in response to glycosylation occupancy changes in Neisseria gonorrhoeae

• DOI: 10.1002/pmic.202300496
• Publication Date: 2024-02-16T04:39:53Z
• Authors: Chris Hadjineophytou; Edmund Loh; Michael Koomey; Nichollas E. Scott
• Keywords: Neisseria gonorrhoeae; Glycoproteomics; Proteome; N-linked glycosylation

Abstract Protein glycosylation is increasingly recognized as a common protein modification across bacterial species. Within the Neisseria genus O ‐linked conserved yet closely related species express O‐ oligosaccharyltransferases (PglOs) with distinct targeting activities. this work, we explore capacity of different PglOs using Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS) fractionation and Data‐Independent Acquisition (DIA) to allow characterization impact changes in on proteome gonorrhoeae . We demonstrate FAIMS expands known glycoproteome wild type N. MS11 enables differences be assessed strains expressing pglO allelic chimeras unique substrate Combining glycoproteomic insights DIA proteomics, that alterations within alleles have widespread impacts Examination peptides targeted by analysis supports occupancy occurs independently levels generally low most glycoproteins. This work thus our understanding roles variation may play governing genus‐level glycosylation.