PXD019548 is an
original dataset announced via ProteomeXchange.
Dataset Summary
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 |
Dataset History
Revision | Datetime | Status | ChangeLog Entry |
0 | 2020-06-03 15:00:28 | ID requested | |
⏵ 1 | 2020-10-13 23:39:03 | announced | |
Publication List
Dataset with its publication pending |
Keyword List
submitter keyword: Methanothermococcus thermolithotrophicus, Archaellin, Glycan, PTM, RPLC-MS, HILIC-MS, NMR |
Contact List
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 | |
Full Dataset Link List
Dataset FTP location
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PRIDE project URI |
Repository Record List
[ + ]
[ - ]
- PRIDE
- PXD019548
- Label: PRIDE project
- Name: Identification of a Novel Glycan N-linked to the Archaellins and S-layer protein of the thermophilic methanogen, Methanothermococcus thermolithotrophicus