Molecular mechanisms regulating the differential association of kainate receptor subunits with SAP90/PSD-95 and SAP97.
Recent studies have demonstrated that kainate receptors are associated with members of the SAP90/PSD-95 family (synapse-associated proteins (SAPs)) in neurons and that SAP90 can cluster and modify the electrophysiological properties of GluR6/KA2 kainate receptors when co-expressed in transfected cells. In vivo, SAP90 tightly binds kainate receptor subunits, while SAP97 is only weakly associated, suggesting that this glutamate receptor differentially associates with SAP90/PSD-95 family members. Here, green fluorescent protein (GFP)-tagged chimeras and deletion mutants of SAP97 and SAP90 were employed to define the molecular mechanism underlying their differential association with kainate receptors. Our results show that a weak interaction between GluR6 and the PDZ1 domain of SAP97 can account for the weak association of GluR6 with the full-length SAP97 observed in vivo. Expression studies in HEK293 cells and in vitro binding studies further show that although the individual Src homology 3 and guanylate kinase domains in SAP97 can interact with the C-terminal tail of KA2 subunit, specific intramolecular interactions in SAP97 (e.g. the SAP97 N terminus (S97N) binding to the Src homology 3 domain) interfere with KA2 binding to the full-length molecule. Because receptor subunits are known to segregate to different parts of the neuron, our results imply that differential association of kainate receptors with SAP family proteins may be one mechanism of subcellular localization.
Department Of Molecular Pharmacology, Physiology, and Biotechnology, Brown University, Providence, Rhode Island 02912, USA.
S
S
Mehta
H
H
Wu
C C
CC
Garner
J
J
Marshall
D048868:Adaptor Proteins, Signal Transducing; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002352:Carrier Proteins / Q000378:metabolism; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D003001:Cloning, Molecular; D000074288:Discs Large Homolog 1 Protein; D006801:Humans; D008565:Membrane Proteins; D008958:Models, Molecular; D009419:Nerve Tissue Proteins / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D021122:Protein Subunits; D051381:Rats; D018092:Kainic Acid Receptors / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism*; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D000074407:SAP90-PSD95 Associated Proteins; D014162:Transfection; D018909:src Homology Domains; D000097806:GluK2 Kainate Receptor
D048868:Adaptor Proteins, Signal Transducing; D002352:Carrier Proteins; D000074288:Discs Large Homolog 1 Protein; D008565:Membrane Proteins; D009419:Nerve Tissue Proteins; D021122:Protein Subunits; D018092:Kainic Acid Receptors; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000074407:SAP90-PSD95 Associated Proteins; D000097806:GluK2 Kainate Receptor; C513314:DLG1 protein, human; C501119:Dlg1 protein, rat