<< Full experiment listing


DataSet Summary

  • HostingRepository: PRIDE
  • AnnounceDate: 2017-10-12
  • AnnouncementXML: Submission_2017-10-12_02:06:48.xml
  • DigitalObjectIdentifier:
  • ReviewLevel: Peer-reviewed dataset
  • DatasetOrigin: Original data
  • RepositorySupport: Unsupported dataset by repository
  • PrimarySubmitter: Kristina Poljak
  • Title: Quantitative Profiling of N-linked Glycosylation Machinery in Yeast Saccharomyces cerevisiae
  • Description: Asparagine-linked glycosylation is a common posttranslational protein modification regulating the structure, stability and function of many proteins. The N-linked glycosylation machinery involves enzymes responsible for the assembly of the lipid-linked oligosaccharide (LLO), which is then transferred to the asparagine residues on the polypeptides by the enzyme oligosaccharyltransferase (OST). A major goal in the study of protein glycosylation is to establish quantitative methods for the analysis of site-specific extent of glycosylation. We developed a sensitive approach to examine glycosylation site occupancy in Saccharomyces cerevisiae by coupling stable isotope labelling (SILAC) approach to parallel reaction monitoring (PRM) mass spectrometry (MS). We combined the method with genetic tools and validated the approach with the identification of novel glycosylation sites dependent on the Ost3p and Ost6p regulatory subunits of OST. Based on the observations that alternations in LLO substrate structure and OST subunits activity differentially alter the systemic output of OST, we conclude that sequon recognition is a direct property of the catalytic subunit Stt3p, whereas auxiliary subunits such as Ost3p and Ost6p extend the OST substrate range by modulating interfering pathways such as protein folding. In addition, our proteomics approach revealed a novel regulatory network that connects isoprenoid lipid biosynthesis and LLO substrate assembly.
  • SpeciesList: scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
  • ModificationList: iodoacetamide derivatized residue
  • Instrument: Q Exactive

Dataset History

VersionDatetimeStatusChangeLog Entry
02017-05-03 02:29:11ID requested
12017-10-12 02:06:49announced

Publication List

  1. Poljak K, Selevsek N, Ngwa E, Grossmann J, Losfeld ME, Aebi M, . Mol Cell Proteomics, 17(1):18-30(2018) [pubmed]

Keyword List

  1. curator keyword: Biological
  2. submitter keyword: ER (Endoplasmatic Reticulum) OST (Oligosaccharyltransferase) GlcNAc (N-acetyl-glucosamine) LLO (Lipid-linked oligosaccharide) MS (Mass spectrometry) PRM (Parallel reaction monitoring) SILAC (Stable isotope labelling with amino acids in cell culture)

Contact List

    Markus Aebi
    • contact affiliation: Institut für Mikrobiologie ETH Zürich, HCI F 407 Vladimir-Prelog-Weg 1-5/10 8093 Zurich Switzerland
    • contact email: markus.aebi@micro.biol.ethz.ch
    • lab head:
    Kristina Poljak
    • contact affiliation: ETH Zurich
    • contact email: poljakk@ethz.ch
    • dataset submitter:

Full Dataset Link List

  1. Dataset FTP location
  2. PRIDE project URI
Repository Record List
Subscribe to receive all new ProteomeXchange announcements!
If you have a question or comment about ProteomeXchange, please contact us!