SNAP-25/syntaxin 1A complex functionally modulates neurotransmitter gamma-aminobutyric acid reuptake.
Neurotransmitter gamma-aminobutyric acid (GABA) release to the synaptic clefts is mediated by the formation of a soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex, which includes two target SNAREs syntaxin 1A and SNAP-25 and one vesicle SNARE VAMP-2. The target SNAREs syntaxin 1A and SNAP-25 form a heterodimer, the putative intermediate of the SNARE complex. Neurotransmitter GABA clearance from synaptic clefts is carried out by the reuptake function of its transporters to terminate the postsynaptic signaling. Syntaxin 1A directly binds to the neuronal GABA transporter GAT-1 and inhibits its reuptake function. However, whether other SNARE proteins or SNARE complex regulates GABA reuptake remains unknown. Here we demonstrate that SNAP-25 efficiently inhibits GAT-1 reuptake function in the presence of syntaxin 1A. This inhibition depends on SNAP-25/syntaxin 1A complex formation. The H3 domain of syntaxin 1A is identified as the binding sites for both SNAP-25 and GAT-1. SNAP-25 binding to syntaxin 1A greatly potentiates the physical interaction of syntaxin 1A with GAT-1 and significantly enhances the syntaxin 1A-mediated inhibition of GAT-1 reuptake function. Furthermore, nitric oxide, which promotes SNAP-25 binding to syntaxin 1A to form the SNARE complex, also potentiates the interaction of syntaxin 1A with GAT-1 and suppresses GABA reuptake by GAT-1. Thus our findings delineate a further molecular mechanism for the regulation of GABA reuptake by a target SNARE complex and suggest a direct coordination between GABA release and reuptake.
Fan
Hua-Ping
HP
Laboratory of Molecular Cell Biology, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China.
Fan
Feng-Juan
FJ
Bao
Lan
L
Pei
Gang
G
D000818:Animals; D050485:GABA Plasma Membrane Transport Proteins / Q000502:physiology*; D058805:GABA Uptake Inhibitors; D009569:Nitric Oxide / Q000502:physiology; D016716:PC12 Cells; D051381:Rats; D017207:Rats, Sprague-Dawley; D050600:SNARE Proteins / Q000502:physiology; D009435:Synaptic Transmission; D050825:Synaptosomal-Associated Protein 25 / Q000737:chemistry / Q000502:physiology*; D050827:Syntaxin 1 / Q000737:chemistry / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000378:metabolism*
D050485:GABA Plasma Membrane Transport Proteins; D058805:GABA Uptake Inhibitors; D050600:SNARE Proteins; C493415:Slc6a1 protein, rat; C494210:Snap25 protein, rat; C494213:Stx1a protein, rat; D050825:Synaptosomal-Associated Protein 25; D050827:Syntaxin 1; D009569:Nitric Oxide; D005680:gamma-Aminobutyric Acid