PXD019548

PXD019548 is an original dataset announced via ProteomeXchange.

Title	Identification of a Novel Glycan N-linked to the Archaellins and S-layer protein of the thermophilic methanogen, Methanothermococcus thermolithotrophicus
Description	Motility in the Archaea domain is facilitated by a unique motility structure termed the archaellum. N-glycosylation of the major structural proteins (archaellins) is important for their subsequent incorporation into the archaellum filament. Here, we report the structure of the archaellin glycan from Methanothermococcus thermolithotrophicus, a methanogen which grows optimally at 65°C. Four archaellin genes (flaB1-4) have previously been identified. In gel digestion and LC-MS analysis revealed the identity of the upper band as FlaB1 and the lower band as FlaB3. Examination of the protein sequences for the four archaellins indicated multiple possible N-linked glycosylation sites in each. We observed using mass spectrometry that Mtc. thermolithotrophicus archaellins is posttranslationally modified at multiple sites with an N-linked branched oligosaccharide composed of 7 sugars (1414 Da). NMR analysis of the purified glycan determined the structure to be α-D-glycero-D-manno-Hep3OMe6OMe-(1-3)-[α-GalNAcA3OMe-(1-2)-]-β-Man-(1-4)-[-GalA3OMe4OAc6CMe-(1-4)--GalA-(1-2)-]-α-GalAN-(1-3)-β-GalNAc-Asn. A detailed investigation by HILIC-MS discovered the presence of several, less abundant glycan variants, related to but distinct from the main heptameric glycan. In addition, we confirmed that the S-layer protein is modified with the same heptameric glycan suggesting a common N-glycosylation pathway.
HostingRepository	PRIDE
AnnounceDate	2020-10-14
AnnouncementXML	Submission_2020-10-13_23:39:02.xml
DigitalObjectIdentifier	https://dx.doi.org/10.6019/PXD019548
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Supported dataset by repository
PrimarySubmitter	John Kelly
SpeciesList	scientific name: Methanothermococcus thermolithotrophicus DSM 2095; NCBI TaxID: 523845;
ModificationList	Deamidated; Oxidation; monohydroxylated residue; deamidated residue
Instrument	Q-Tof Ultima; Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2020-06-03 15:00:28	ID requested
⏵ 1	2020-10-13 23:39:03	announced

Dataset with its publication pending

submitter keyword: Methanothermococcus thermolithotrophicus, Archaellin, Glycan, PTM, RPLC-MS, HILIC-MS, NMR

John Francis Kelly
contact affiliation	Human Health Therapeutics, National Research Council of Canada
contact email	John.Kelly@nrc-cnrc.gc.ca
lab head
John Kelly
contact affiliation	Human Health Therapeutics, National Research Council of Canada
contact email	John.Kelly@nrc-cnrc.gc.ca
dataset submitter

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PRIDE project URI

• PRIDE
  1. PXD019548
     1. Label: PRIDE project
     2. Name: Identification of a Novel Glycan N-linked to the Archaellins and S-layer protein of the thermophilic methanogen, Methanothermococcus thermolithotrophicus