SAP97 blocks the RXR ER retention signal of NMDA receptor subunit GluN1-3 through its SH3 domain

Biochim Biophys Acta. 2015 Feb;1853(2):489-99. doi: 10.1016/j.bbamcr.2014.11.030. Epub 2014 Dec 9.

Abstract

SAP97 is directly involved in exporting NMDA receptors with a specific subunit composition from the endoplasmic reticulum (ER). Characterization of the interactions between SAP97 and an NMDA receptor splice variant, GluN1-3, and of the effects on forward trafficking revealed that an ER-level interaction blocked the RXR ER-retention motif in the GluN1-3 cytoplasmic C-terminus in the context of both reporter molecules and full-length receptors. Binding of SAP97 to the PDZ-binding domain of GluN1-3 was required, but the blockade of ER-retention was mediated by the SH3-GuK domains coupled with the action of the N-terminus of SAP97. While other domains of SAP97 were involved in forward trafficking of GluN1-3 out of the ER, the SH3 domain was necessary and sufficient to block the ER retention. This is the first direct evidence for the masking of ER-retention signals by PDZ domain-containing proteins, and provides detailed underlying mechanistic requirements. Such a mechanism could be central to modulating the ER exit of receptors into local, non-conventional or conventional, secretory pathways in neurons.

Keywords: NMDA; PDZ domain; RXR motif; Receptor; SAP97; SH3 domain; Trafficking.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adaptor Proteins, Signal Transducing / chemistry
  • Adaptor Proteins, Signal Transducing / metabolism*
  • Amino Acid Sequence
  • Animals
  • COS Cells
  • Cell Membrane / metabolism
  • Chlorocebus aethiops
  • Discs Large Homolog 1 Protein
  • Endoplasmic Reticulum / metabolism*
  • Green Fluorescent Proteins / metabolism
  • HeLa Cells
  • Humans
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism*
  • Models, Biological
  • Molecular Sequence Data
  • Protein Subunits / chemistry
  • Protein Subunits / metabolism*
  • Receptors, N-Methyl-D-Aspartate / chemistry*
  • Receptors, N-Methyl-D-Aspartate / metabolism*
  • Retinoid X Receptors / metabolism*
  • Structure-Activity Relationship
  • src Homology Domains*

Substances

  • Adaptor Proteins, Signal Transducing
  • DLG1 protein, human
  • Discs Large Homolog 1 Protein
  • Membrane Proteins
  • Protein Subunits
  • Receptors, N-Methyl-D-Aspartate
  • Retinoid X Receptors
  • Green Fluorescent Proteins