PXD017151

PXD017151 is an original dataset announced via ProteomeXchange.

Title	Glycine receptor complex analysis using immunoprecipitation-blue native gel electrophoresis-mass spectrometry
Description	The pentameric glycine receptor (GlyR), comprising the α1 and β subunits, is a major inhibitory ionotropic receptor in brainstem and spinal cord. GlyRs interact with gephyrin (GPHN), a scaffold protein that anchors the GlyR in the plasma membrane and enables it to form clusters in glycinergic postsynapses. Using an interaction proteomics approach, we provides evidence of the ArfGEFs IQ motif and Sec7 domain 3 (IQSEC3) and IQ motif and Sec7 domain 2 (IQSEC2) as two novel synaptic proteins interacting with GlyR complexes. When the affinity-isolated GlyR complexes were fractionated by blue native gel electrophoresis and characterized by mass spectrometry, GlyR α1β-GPHN appeared as the most abundant complex with a molecular weight of approximately 1 MDa, and GlyR α1β-GPHN-IQSEC3 as a minor protein complex of approximately 1.2 MDa. A third GlyR α1β-GPHN-IQSEC2 complex existed at the lowest amount with a mass similar to the IQSEC3-containing complex. Using yeast two-hybrid we demonstrate that IQSEC3 interacts with the GlyR complex by binding to the GPHN G domain at the N-terminal of the IQSEC3 IQ-like domain. Our data provide direct evidence of the interaction of IQSEC3 with GlyR-GPHN complexes, underscoring a potential role of these ArfGEFs in the function of glycinergic synapses.
HostingRepository	PRIDE
AnnounceDate	2023-11-14
AnnouncementXML	Submission_2023-11-14_07:40:35.705.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Sophie van der Spek
SpeciesList	scientific name: Mus musculus (Mouse); NCBI TaxID: 10090;
ModificationList	No PTMs are included in the dataset
Instrument	TripleTOF 5600

Revision	Datetime	Status	ChangeLog Entry
0	2020-01-16 05:51:14	ID requested
1	2022-12-12 08:17:10	announced
⏵ 2	2023-11-14 07:40:39	announced	2023-11-14: Updated project metadata.

van der Spek SJF, Koopmans F, Paliukhovich I, Ramsden SL, Harvey K, Harvey RJ, Smit AB, Li KW, Glycine Receptor Complex Analysis Using Immunoprecipitation-Blue Native Gel Electrophoresis-Mass Spectrometry. Proteomics, 20(3-4):e1900403(2020) [pubmed]

submitter keyword: blue native gel electrophoresis, proteomics, protein complex, glycine receptor, IQSEC3

Ka Wan Li
contact affiliation	MCN, Neuroproteomics, CNCR, VU, Amsterdam
contact email	k.w.li@vu.nl
lab head
Sophie van der Spek
contact affiliation	CNCR, VU Amsterdam
contact email	sophiejf@hotmail.com
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/2022/12/PXD017151

PRIDE project URI

• PRIDE
  1. PXD017151
     1. Label: PRIDE project
     2. Name: Glycine receptor complex analysis using immunoprecipitation-blue native gel electrophoresis-mass spectrometry