Direct interaction of N-ethylmaleimide-sensitive factor with GABAA receptor β subunits

• DOI: 10.1016/j.mcn.2005.07.006
• Publication Date: 2005-08-11T13:12:48Z
• Authors: Hidefumi Goto; Miho Terunuma; Takashi Kanematsu; Yoshio Misumi; Stephen J. Moss; Masato Hirata
• Keywords: 0303 health sciences; Base Sequence; Recombinant Fusion Proteins; Molecular Sequence Data; Vesicular Transport Proteins; Kidney; Receptors, GABA-A; Peptide Fragments; Cell Line; Mice; 03 medical and health sciences; Receptors, GABA; COS Cells; Chlorocebus aethiops; Mutagenesis, Site-Directed; Animals; Humans; Amino Acid Sequence; N-Ethylmaleimide-Sensitive Proteins

GABA(A) receptors mediate most of the fast inhibitory neurotransmission in the brain, and are believed to be composed mainly of alpha, beta, and gamma subunits. It has been shown that GABA(A) receptors interact with a number of binding partners that act to regulate both receptor function and cell surface stability. Here, we reveal that GABA(A) receptors interact directly with N-ethylmaleimide-sensitive factor (NSF), a critical regulator of vesicular dependent protein trafficking, as measured by in vitro protein binding and co-immunoprecipitation assays. In addition, we established that NSF interacts with residues 395-415 of the receptor beta subunits and co-localizes with GABA(A) receptors in hippocampal neurons. We also established that NSF can regulate GABA(A) receptor cell surface expression depending upon residues 395-415 in the beta3 subunit. Together, our results suggest an important role for NSF activity in regulating the cell surface stability of GABA(A) receptors.