Identification of PIST PDZ domain interactors

Jim Y. Chen*, Dane M Chetkovich

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

Glutamate receptors are proteins that mediate most excitatory communication between neurons in the brain. One type of glutamate receptor, alpha-amino-3-hydroxy-5-methylisoxazolepropionate receptor (AMPAR), is moved to and from the synapse to control the strength of synaptic transmission. This movement is critical for the synaptic changes that underlie some forms of learning and memory. Abnormal targeting of AMPARs can cause epilepsy and learning disorders. Proteins interacting specifically with TC10 (PIST) are important for AMPAR targeting synapses; it acts as a chaperone for the AMPAR and an AMPAR-binding protein, stargazin. PIST contains several important domains, regions that are conserved between many proteins, and are known to have specific functions. For PIST, these include two coiled-coil domains, a leucine zipper, a PDZ domain (named after several proteins that contain this domain, PSD-95, Discs Large, and Zona Occludens), and an acid cluster. PIST interacts with several proteins through its coiled-coil region, and its PDZ domain is known to bind to and traffic many transmembrane proteins outside the brain. PIST binds to stargazin/AMPAR at amino acid residues separate from its named domains. However, experiments in our lab suggest that, although the PIST-PDZ domain does not bind stargazin/AMPA receptors, it is also critical for the synaptic targeting of these proteins. Because the PIST-PDZ domain is important for AMPAR synaptic targeting, we sought to identify novel proteins that interact with the PIST-PDZ domain. Using an assay of protein-protein interaction, the yeast two-hybrid system, we screened a cDNA library of brain proteins using the PIST-PDZ domain and obtained 100 possible interacting clones. By reintroducing these clones into yeast containing the PIST-PDZ domain, we sought to confirm specific interaction with PIST. We performed restriction digest and DNA sequencing of interacting clones to identify the interacting proteins. By identifying PIST-interacting proteins, we wanted to increase knowledge on the mechanism by which PIST serves as a chaperone for stargazin/AMPAR in their delivery to the synapse.

Original languageEnglish (US)
JournalEthnicity and Disease
Volume15
Issue number3 SUPPL. 4
StatePublished - Jun 1 2005

ASJC Scopus subject areas

  • Epidemiology

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