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Purification of the Dendritic Filopodia-rich Fraction
Published on: May 2, 2019
Neto1 associates with the NMDA receptor/amyloid precursor protein complex
Sarah L Cousins1, Neal Innocent, F Anne Stephenson
1University College London School of Pharmacy, London, UK.
Journal of Neurochemistry
|April 30, 2013
Summary
Neuropilin tolloid-like 1 (Neto1) is a novel component of NMDA receptor complexes. This study found Neto1 associates with amyloid precursor protein (APP) and NMDA receptors in brain tissue.
Area of Science:
- Neuroscience
- Molecular Biology
- Cell Biology
Background:
- Neuropilin tolloid-like 1 (Neto1) is a transmembrane protein identified as a novel component of NMDA receptor complexes.
- Previous work identified NMDA receptor trafficking complexes involving amyloid precursor protein (APP)695/GluN1/GluN2A and APP695/GluN1/GluN2B.
Purpose of the Study:
- To investigate the association of Neto1 with APP/NMDA receptor trafficking complexes.
- To determine Neto1's role in the surface expression of these complexes.
Main Methods:
- Co-immunoprecipitation assays using mammalian cells and native brain tissue.
- Transfection of cells with APP695(FLAG), Neto1(HA), and NMDA receptor subunits (GluN1/GluN2A or GluN1/GluN2B).
- Mapping of the Neto1 binding domain on GluN2A using chimeras and truncation constructs.
Main Results:
- Neto1 co-immunoprecipitated with NMDA receptors (via GluN2A/GluN2B) but not directly with APP695.
- APP, NMDA receptors, and Neto1 co-exist within a single macromolecular complex in both cell lines and brain tissue.
- Neto1 reduced the surface expression of NMDA receptors but did not affect APP or PSD-95 enhanced surface expression.
- Neto1 binding to GluN2A involves its intracellular tail, and this interaction is linked to the association with SAP102.
Conclusions:
- Neto1 is a component of APP/NMDA receptor trafficking complexes.
- Neto1 influences NMDA receptor surface expression, potentially through interactions mediated by the intracellular tail of GluN2A and SAP102.
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