title: Specific interaction of postsynaptic densities with membrane rafts isolated from synaptic plasma membranes.
abstract: Postsynaptic membrane rafts are believed to play important roles in synaptic signaling, plasticity, and maintenance. We recently demonstrated the presence, at the electron microscopic level, of complexes consisting of membrane rafts and postsynaptic densities (PSDs) in detergent-resistant membranes (DRMs) prepared from synaptic plasma membranes (SPMs) ( Suzuki et al., 2011 , J Neurochem, 119, 64-77). To further explore these complexes, here we investigated the nature of the binding between purified SPM-DRMs and PSDs in vitro. In binding experiments, we used SPM-DRMs prepared after treating SPMs with n-octyl-β-d-glucoside, because at concentrations of 1.0% or higher it completely separates SPM-DRMs and PSDs, providing substantially PSD-free unique SPM-DRMs as well as DRM-free PSDs. PSD binding to PSD-free DRMs was identified by mass spectrometry, Western blotting, and electron microscopy. PSD proteins were not incorporated into SPMs, and significantly less PSD proteins were incorporated into DRMs prepared from liver membranes, providing in vitro evidence that binding of PSDs to DRMs is specific and suggestion of the presence of specific interacting molecules. These specific interactions may have important roles in synaptic development, function, and plasticity in vivo. In addition, the binding system we developed may be a good tool to search for binding molecules and binding mechanisms between PSDs and rafts.
authors.affiliation: Department of Neuroplasticity, Institute of Pathogenesis and Disease Prevention, Shinshu University Graduate School of Medicine, Matsumoto, Japan.
authors.forename: Qian
authors.initials: Q
authors.lastname: Liu
authors.forename: Wei-Dong
authors.initials: WD
authors.lastname: Yao
authors.forename: Tatsuo
authors.initials: T
authors.lastname: Suzuki
mesh_terms: D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D001921:Brain / Q000166:cytology / Q000648:ultrastructure; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2 / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism; D005677:G(M1) Ganglioside / Q000378:metabolism; D008297:Male; D021962:Membrane Microdomains / Q000378:metabolism* / Q000648:ultrastructure; D046529:Microscopy, Electron, Transmission; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D057907:Post-Synaptic Density / Q000378:metabolism* / Q000648:ultrastructure; D051381:Rats; D017208:Rats, Wistar; D018091:Receptors, AMPA / Q000378:metabolism; D013570:Synaptic Membranes / Q000378:metabolism* / Q000648:ultrastructure
chemical_list: D009419:Nerve Tissue Proteins; D018091:Receptors, AMPA; D005677:G(M1) Ganglioside; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D002784:Cholesterol; D054732:Calcium-Calmodulin-Dependent Protein Kinase Type 2; C104722:glutamate receptor ionotropic, AMPA 2