The PDZ domains of postsynaptic density (PSD) protein-95 play a role in the localization of PSD-95 and binding partners to neuronal synapses. The identification of binding partners to these PDZ domains can help us in understanding how signalling complexes are assembled. We observed that one of the subunits in the sec6/8 or exocyst complex, sec8, contains a C-terminal consensus sequence for PDZ binding. Sec8 binds to PDZ1–2 of PSD-95, and this binding can be competed with a peptide that binds to PDZ1 and PDZ2 in the peptide-binding site. In addition, binding of sec8 is dependent on its C-terminal-binding sequence namely Thr-Thr-Val (TTV). Immunoblotting of rat tissue extracts shows that sec8 and PSD-95 are enriched in the same brain regions, and sec8 and PSD-95 have the same subcellular distribution in pheochromocytoma cells, suggesting that these proteins may interact in vivo. Immunoprecipitation studies of sec8 and PSD-95 in brain provide further evidence of a sec8 and PSD-95 interaction. Furthermore, the cytosolic PSD-95 interactor competes with sec8 for interaction with PSD-95. Taken together, our results suggest that the cytosolic PSD-95 interactor may function to regulate the ability of sec8 to bind to PSD-95.
- postsynaptic density
- protein targeting
↵1 These authors contributed equally to this work.
Abbreviations used: PSD, postsynaptic density; cypin, cytosolic PSD-95 interactor; DIV, days in vitro; DLG, discs large; GFP, green fluorescent protein; GK, guanylate kinase; GST, glutathione S-transferase; MAGUK, membrane-associated guanylate kinase; MTOC, microtubule organizing centre; NGF, nerve growth factor; NMDA, N-methyl-d-aspartate; nNOS, neuronal nitric oxide synthase; PC12, pheochromocytoma; SAP, synapse-associated protein; SH3 domain, src homology 3 domain.
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