Glycine Receptor Complex Analysis Using Immunoprecipitation‐Blue Native Gel Electrophoresis‐Mass Spectrometry
Abstract: 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, 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 is provided. When the affinity‐isolated GlyR complexes are fractionated by blue native gel electrophoresis and characterized by mass spectrometry, GlyR α1β‐GPHN appears as the most abundant complex with a molecular weight of ≈1 MDa, and GlyR α1β‐GPHN‐IQSEC3 as a minor protein complex of ≈1.2 MDa. A third GlyR α1β‐GPHN‐IQSEC2 complex exists at the lowest amount with a mass similar to the IQSEC3 containing complex. Using yeast two‐hybrid it is demonstrated that IQSEC3 interacts with the GlyR complex by binding to the GPHN G domain at the N‐terminal of the IQSEC3 IQ‐like domain. The 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.
- Standort
-
Deutsche Nationalbibliothek Frankfurt am Main
- Umfang
-
Online-Ressource
- Sprache
-
Englisch
- Erschienen in
-
Glycine Receptor Complex Analysis Using Immunoprecipitation‐Blue Native Gel Electrophoresis‐Mass Spectrometry ; volume:20 ; number:3-4 ; year:2020 ; extent:9
Proteomics ; 20, Heft 3-4 (2020) (gesamt 9)
- Urheber
-
van der Spek, Sophie J. F.
Koopmans, Frank
Paliukhovich, Iryna
Ramsden, Sarah L.
Harvey, Kirsten
Harvey, Robert J.
Smit, August B.
Li, Ka Wan
- DOI
-
10.1002/pmic.201900403
- URN
-
urn:nbn:de:101:1-2022061708311682336257
- Rechteinformation
-
Open Access; Der Zugriff auf das Objekt ist unbeschränkt möglich.
- Letzte Aktualisierung
-
15.08.2025, 07:39 MESZ
Datenpartner
Deutsche Nationalbibliothek. Bei Fragen zum Objekt wenden Sie sich bitte an den Datenpartner.
Beteiligte
- van der Spek, Sophie J. F.
- Koopmans, Frank
- Paliukhovich, Iryna
- Ramsden, Sarah L.
- Harvey, Kirsten
- Harvey, Robert J.
- Smit, August B.
- Li, Ka Wan
Ähnliche Objekte (12)
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Analysis of protein complexes in Plasmodium falciparum : using blue native polyacrylamide gel electrophoresis and label-free quantitative mass spectrometry
Ocular Proteomics with Emphasis on Two-Dimensional Gel Electrophoresis and Mass Spectrometry
Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis
Capillary electrophoresis-, liquid chromatography-mass spectrometry
The practice of quantitative gel electrophoresis
Capillary electrophoresis‐mass spectrometry for protein analyses under native conditions: Current progress and perspectives
Useful Buffer and Gel Systems for Polyacrylamide Gel Electrophoresis
Two-dimensional blue native/blue native polyacrylamide gel electrophoresis for the characterization of mitochondrial protein complexes and supercomplexes.
Two-Dimensional Gel Electrophoresis, Isoelectric Focusing and Agarose Gel Electrophoresis in the Diagnosis of Multiple Sclerosis
Proteomic profiling of the mesenteric lymph after hemorrhagic shock: Differential gel electrophoresis and mass spectrometry analysis
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Analysis of protein complexes in Plasmodium falciparum : using blue native polyacrylamide gel electrophoresis and label-free quantitative mass spectrometry
Ocular Proteomics with Emphasis on Two-Dimensional Gel Electrophoresis and Mass Spectrometry
Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis
Capillary electrophoresis-, liquid chromatography-mass spectrometry
The practice of quantitative gel electrophoresis
Capillary electrophoresis‐mass spectrometry for protein analyses under native conditions: Current progress and perspectives
Useful Buffer and Gel Systems for Polyacrylamide Gel Electrophoresis
Two-dimensional blue native/blue native polyacrylamide gel electrophoresis for the characterization of mitochondrial protein complexes and supercomplexes.
Two-Dimensional Gel Electrophoresis, Isoelectric Focusing and Agarose Gel Electrophoresis in the Diagnosis of Multiple Sclerosis
Proteomic profiling of the mesenteric lymph after hemorrhagic shock: Differential gel electrophoresis and mass spectrometry analysis
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Native Proteomics by Capillary Zone Electrophoresis‐Mass Spectrometry
Analysis of protein complexes in Plasmodium falciparum : using blue native polyacrylamide gel electrophoresis and label-free quantitative mass spectrometry
Ocular Proteomics with Emphasis on Two-Dimensional Gel Electrophoresis and Mass Spectrometry
Disc electrophoresis and related techniques of polyacrylamide gel electrophoresis
Capillary electrophoresis-, liquid chromatography-mass spectrometry
The practice of quantitative gel electrophoresis
Capillary electrophoresis‐mass spectrometry for protein analyses under native conditions: Current progress and perspectives
Useful Buffer and Gel Systems for Polyacrylamide Gel Electrophoresis
Two-dimensional blue native/blue native polyacrylamide gel electrophoresis for the characterization of mitochondrial protein complexes and supercomplexes.
Two-Dimensional Gel Electrophoresis, Isoelectric Focusing and Agarose Gel Electrophoresis in the Diagnosis of Multiple Sclerosis