Virulence of the gastric pathogen Helicobacter pylori (Hp) is directly linked to the pathogen's ability to glycosylate proteins. Although Hp's glycans are linked to pathogenesis, Hp's glycome remains poorly understood. Here we set out to facilitate the discovery of Hp's glycoproteins by coupling glycan metabolic labeling with mass spectrometry analysis. An assessment of the subcellular distribution of azide-labeled proteins by Western blot analysis indicated that glycoproteins are present throughout Hp and may therefore serve diverse functions. To identify these species, Hp's azide-labeled glycoproteins were tagged via Staudinger ligation, enriched by tandem affinity chromatography, and analyzed by multidimensional protein identification technology. We identified 125 candidate glycoproteins with diverse biological functions, including known links to pathogenesis. Of these, the secreted virulence factor urease was biochemically validated as glycosylated. Broadly, this report validates metabolic labeling as an effective and efficient approach for the identification and study of bacterial glycoproteins.

Targeted identification of glycosylated proteins in the gastric pathogen <i>Helicobacter pylori</i>

245th National Meeting (2013)

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Targeted identification of glycosylated proteins in the gastric pathogen Helicobacter pylori