Proteomic Analysis of an Interactome for Long-Form AMPA Receptor Subunits
journal contributionposted on 2010-04-05, 00:00 authored by Sandra D. Santos, Bruno Manadas, Carlos B. Duarte, Ana Luísa Carvalho
Glutamate receptors of the AMPA-type mediate fast excitatory synaptic transmission in the central nervous system and play key roles in synaptic plasticity. The binding of these receptors to a variety of proteins is known to regulate their targeting to the synapse and consequently to modulate synaptic strength, as well as to modify receptor characteristics. In this study, a proteomic screening was conducted in order to identify new binding partners for GluR4 AMPA receptor subunit. Immunoprecipitation of GluR4 and associated proteins was performed using rat cerebellum lysates and an heterologous systems overexpressing GluR4 AMPA receptor subunit. Isolated immuno-complexes were resolved by 1-D SDS-PAGE, and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). This approach led to the identification of several interactors, most of which are novel AMPA receptor partners, namely, cytoskeleton proteins, motor proteins, RNA processing proteins which are part of neuronal RNA granules, and kinases, among others. This study unravels new constituents of the macromolecular complex of long-form calcium-permeable AMPA receptors.
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RNA granulesRNA processing proteinsrat cerebellum lysatesbinding partnersheterologous systems overexpressing GluR 4 AMPA receptor subunitmotor proteinsproteomic screeningreceptor characteristicsProteomic AnalysisGluR 4 AMPA receptor subunitstudy unravelsnovel AMPA receptor partnerschromatography tandem mass spectrometrysynaptic strengthGluR 4cytoskeleton proteinssynaptic plasticitysynaptic transmission