PXD006435

PXD006435 is an original dataset announced via ProteomeXchange.

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.
HostingRepository	PRIDE
AnnounceDate	2017-10-12
AnnouncementXML	Submission_2017-10-12_02:06:48.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Kristina Poljak
SpeciesList	scientific name: Saccharomyces cerevisiae (Baker's yeast); NCBI TaxID: 4932;
ModificationList	iodoacetamide derivatized residue
Instrument	Q Exactive

Revision	Datetime	Status	ChangeLog Entry
0	2017-05-03 02:29:11	ID requested
⏵ 1	2017-10-12 02:06:49	announced

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

curator keyword: Biological

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)

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

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2017/10/PXD006435

PRIDE project URI

• PRIDE
  1. PXD006435
     1. Label: PRIDE project
     2. Name: Quantitative Profiling of N-linked Glycosylation Machinery in Yeast Saccharomyces cerevisiae