title: Neto1 associates with the NMDA receptor/amyloid precursor protein complex.
abstract: Neuropilin tolloid-like 1 (Neto1), is a CUB domain-containing transmembrane protein that was recently identified as a novel component of the NMDA receptor complex. Here, we have investigated the possible association of Neto1 with the amyloid precursor protein (APP)695/GluN1/GluN2A and APP695/GluN1/GluN2B NMDA receptor trafficking complexes that we have previously identified. Neto1(HA) was shown to co-immunoprecipitate with assembled NMDA receptors via GluN2A or GluN2B subunits; Neto1(HA) did not co-immunoprecipitate APP695(FLAG) . Co-immunoprecipitations from mammalian cells co-transfected with APP695(FLAG) , Neto1(HA) and GluN1/GluN2A or GluN1/GluN2B revealed that all four proteins co-exist within one macromolecular complex. Immunoprecipitations from native brain tissue similarly revealed the existence of a GluN1/GluN2A or GluN2B/APP/Neto1 complex. Neto1(HA) caused a reduction in the surface expression of both NMDA receptor subtypes, but had no effect on APP695(FLAG) - or PSD-95α(c-Myc) enhanced surface receptor expression. The Neto1 binding domain of GluN2A was mapped using GluN1/GluN2A chimeras and GluN2A truncation constructs. The extracellular GluN2A domain does not contribute to association with Neto1(HA) but deletion of the intracellular tail resulted in a loss of Neto-1(HA) co-immunoprecipitation which was paralleled by a loss of association between GluN2A and SAP102. Thus, Neto1 is concluded to be a component of APP/NMDA receptor trafficking complexes.
authors.affiliation: University College London School of Pharmacy, London, UK.
authors.forename: Sarah L
authors.initials: SL
authors.lastname: Cousins
authors.forename: Neal
authors.initials: N
authors.lastname: Innocent
authors.forename: F Anne
authors.initials: FA
authors.lastname: Stephenson
mesh_terms: D016564:Amyloid beta-Protein Precursor / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D057809:HEK293 Cells; D006801:Humans; D007150:Immunohistochemistry; D047468:Immunoprecipitation; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D050506:Mutant Chimeric Proteins / Q000235:genetics / Q000378:metabolism; D009419:Nerve Tissue Proteins / Q000235:genetics; D011956:Receptors, Cell Surface / Q000235:genetics / Q000378:metabolism; D016194:Receptors, N-Methyl-D-Aspartate / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D016564:Amyloid beta-Protein Precursor; D008565:Membrane Proteins; D050506:Mutant Chimeric Proteins; D009419:Nerve Tissue Proteins; D011956:Receptors, Cell Surface; D016194:Receptors, N-Methyl-D-Aspartate; C485668:GRIN1 protein, human; C456602:NETO1 protein, human
keywords: Alzheimer's disease; NMDA receptors; Neto1; SAP102; amyloid precursor protein; post-synaptic density 95