title: Functional modulation of the glutamate transporter variant GLT1b by the PDZ domain protein PICK1.
abstract: The dominant glutamate transporter isoform in the mammalian brain, GLT1, exists as at least three splice variants, GLT1a, GLT1b, and GLT1c. GLT1b interacts with the scaffold protein PICK1 (protein interacting with kinase C1), which is implicated in glutamatergic neurotransmission via its regulatory effect on trafficking of AMPA-type glutamate receptors. The 11 extreme C-terminal residues specific for the GLT1b variant are essential for its specific interaction with the PICK1 PDZ domain, but a functional consequence of this interaction has remained unresolved. To identify a functional effect of PICK1 on GLT1a or GLT1b separately, we employed the Xenopus laevis expression system. GLT1a and GLT1b displayed similar electrophysiological properties and EC50 for glutamate. Co-expressed PICK1 localized efficiently to the plasma membrane and resulted in a 5-fold enhancement of the leak current in GLT1b-expressing oocytes with only a minor effect on [(3)H]glutamate uptake. Three different GLT1 substrates all caused a slow TBOA-sensitive decay in the membrane current upon prolonged application, which provides support for the leak current being mediated by GLT1b itself. Leak and glutamate-evoked currents in GLT1a-expressing oocytes were unaffected by PICK1 co-expression. PKC activation down-regulated GLT1a and GLT1b activity to a similar extent, which was not affected by co-expression of PICK1. In conclusion, PICK1 may not only affect glutamatergic neurotransmission by its regulatory effect on glutamate receptors but may also affect neuronal excitability via an increased GLT1b-mediated leak current. This may be particularly relevant in pathological conditions such as amyotrophic lateral sclerosis and cerebral hypoxia, which are associated with neuronal GLT1b up-regulation.
authors.affiliation: Department of Cellular and Molecular Medicine, University of Copenhagen, DK-2200 Copenhagen N, Denmark.
authors.forename: Rikke
authors.initials: R
authors.lastname: Sogaard
authors.forename: Lars
authors.initials: L
authors.lastname: Borre
authors.forename: Thomas H
authors.initials: TH
authors.lastname: Braunstein
authors.forename: Kenneth L
authors.initials: KL
authors.lastname: Madsen
authors.forename: Nanna
authors.initials: N
authors.lastname: MacAulay
mesh_terms: D017398:Alternative Splicing; D000818:Animals; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D003598:Cytoskeletal Proteins; D027342:Excitatory Amino Acid Transporter 2 / Q000235:genetics / Q000378:metabolism*; D005260:Female; D018698:Glutamic Acid / Q000378:metabolism* / Q000494:pharmacology; D008564:Membrane Potentials / Q000187:drug effects / Q000502:physiology; D008856:Microscopy, Fluorescence; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism*; D009865:Oocytes / Q000378:metabolism / Q000502:physiology; D011485:Protein Binding; D020033:Protein Isoforms / Q000235:genetics / Q000378:metabolism; D011493:Protein Kinase C / Q000378:metabolism; D051381:Rats; D014316:Tritium; D014982:Xenopus laevis
chemical_list: D002352:Carrier Proteins; D003598:Cytoskeletal Proteins; D027342:Excitatory Amino Acid Transporter 2; D009687:Nuclear Proteins; C498298:PICK1 protein, rat; D020033:Protein Isoforms; C493395:Slc1a2 protein, rat; D014316:Tritium; D018698:Glutamic Acid; D011493:Protein Kinase C
keywords: Amyotrophic Lateral Sclerosis (Lou Gehrig's Disease); BAR Domain; Glutamate; Leak Current; Neurotransmitter Transport; PDZ Domain; PICK1; Protein Kinase C (PKC); Protein-Protein Interactions