title: Lactobacillus plantarum WCFS1 O-linked protein glycosylation: an extended spectrum of target proteins and modification sites detected by mass spectrometry.
abstract: It has recently been shown that the major autolysin Acm2 from Lactobacillus plantarum WCFS1 undergoes intracellular O-GlcNAcylation [Fredriksen L, Mathiesen G, Moen A, Bron PA, Kleerebezem M, Eijsink VG, Egge-Jacobsen W. 2012. The major autolysin Acm2 from Lactobacillus plantarum undergoes cytoplasmic O-glycosylation. J Bacteriol. 194(2):325-333]. To gain more insight into the occurrence of this protein modification, methods based on the higher energy collisional fragmentation of the Orbitrap XL mass spectrometer to generate both diagnostic oxonium (glycan) ions and significant peptide sequencing information were used to detect and identify novel glycoproteins. This led to the identification of 10 novel glycoproteins, including four proteins with well-known functions in the cytoplasm, a compartment not previously recognized to contain glycosylated proteins in bacteria: the molecular chaperone DnaK, the E2 subunit of the pyruvate dehydrogenase complex PdhC, the signal recognition particle receptor FtsY and the DNA translocase FtsK1. Among the other, glycosylated proteins were two extracellular peptidoglycan hydrolases and a mucus-binding protein. In total, 49 glycosylation sites for N-acetylhexosamine (HexNAc) were detected in the 11 Lactobacillus glycoproteins found so far. Most of the attached glycans consisted of a single HexNAc per site, whereas hexose moieties were also found in a few cases (in both of the peptidoglycan hydrolases and in DnaK).
authors.affiliation: Department of Chemistry, Biotechnology and Food Science, Norwegian University of Life Sciences, 1432 Aas, Norway.
authors.forename: Lasse
authors.initials: L
authors.lastname: Fredriksen
authors.forename: Anders
authors.initials: A
authors.lastname: Moen
authors.forename: Alexei A
authors.initials: AA
authors.lastname: Adzhubei
authors.forename: Geir
authors.initials: G
authors.lastname: Mathiesen
authors.forename: Vincent G H
authors.initials: VG
authors.lastname: Eijsink
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Egge-Jacobsen
mesh_terms: D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry; D006031:Glycosylation; D048191:Lactiplantibacillus plantarum / Q000737:chemistry*; D013058:Mass Spectrometry
chemical_list: D006023:Glycoproteins
keywords: bacterial O-linked glycosylation; glycoproteins; mass spectrometry; post-translational modifications; shotgun proteomics