Substrates regulate gamma-aminobutyric acid transporters in a syntaxin 1A-dependent manner.
Several ion channels and pumps are regulated by syntaxin 1A, a component of the synaptic vesicle docking and fusion apparatus. One such regulated protein is the rat brain gamma-aminobutyric acid (GABA) transporter GAT1. The N-terminal cytoplasmic domain of GAT1 directly interacts with syntaxin 1A; this interaction induces a decrease in the rate at which GABA and associated ions are transported. GAT1 function also is regulated by transporter substrates, raising the possibility that substrates mediate at least some of their effects by regulating the interaction between GAT1 and syntaxin 1A. In oocytes expressing GAT1 and syntaxin 1A, superfusion of transporter substrates increases the GAT1 transport rate. The substrate-induced rate change (i) is prevented by coapplication of GAT1 antagonists, (ii) does not occur in oocytes expressing GAT1 alone, and (iii) does not occur in oocytes expressing interaction-deficient syntaxin 1A mutants. In oocytes, and in hippocampal neurons that endogenously express both GAT1 and syntaxin 1A, substrate application results in a decrease in the fraction of syntaxin 1A that is bound to GAT1 on a time-scale comparable to the substrate-induced change in transport rates. These data suggest that substrate translocation regulates GAT1-syntaxin 1A interactions and provide a mechanism by which GABA transport can be increased during times of rising synaptic GABA concentrations.
Quick
Michael W
MW
Department of Neurobiology, CIRC 446, University of Alabama, 1719 6th Avenue South, Birmingham, AL 35294, USA. quick@nrc.uab.edu
D000818:Animals; D000831:Animals, Newborn; D000954:Antigens, Surface / Q000378:metabolism*; D019921:Biotinylation; D002352:Carrier Proteins / Q000378:metabolism*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004594:Electrophysiology; D050485:GABA Plasma Membrane Transport Proteins; D006624:Hippocampus / Q000166:cytology / Q000378:metabolism; D007477:Ions; D008565:Membrane Proteins / Q000378:metabolism*; D026901:Membrane Transport Proteins; D009154:Mutation; D009419:Nerve Tissue Proteins / Q000378:metabolism*; D009557:Nipecotic Acids / Q000494:pharmacology; D027361:Organic Anion Transporters; D011233:Precipitin Tests; D011485:Protein Binding; D011487:Protein Conformation; D017434:Protein Structure, Tertiary; D051381:Rats; D015398:Signal Transduction; D050827:Syntaxin 1; D013997:Time Factors; D015854:Up-Regulation
D000954:Antigens, Surface; D002352:Carrier Proteins; D050485:GABA Plasma Membrane Transport Proteins; D007477:Ions; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D009557:Nipecotic Acids; D027361:Organic Anion Transporters; C493415:Slc6a1 protein, rat; C494213:Stx1a protein, rat; D050827:Syntaxin 1; C030278:nipecotic acid