title: Direct interaction of N-ethylmaleimide-sensitive factor with GABA(A) receptor beta subunits.
abstract: 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.
authors.affiliation: Laboratory of Molecular and Cellular Biochemistry, Faculty of Dental Science and Station for Collaborative Research, Kyushu University, Fukuoka 812-8582, Japan.
authors.forename: Hidefumi
authors.initials: H
authors.lastname: Goto
authors.forename: Miho
authors.initials: M
authors.lastname: Terunuma
authors.forename: Takashi
authors.initials: T
authors.lastname: Kanematsu
authors.forename: Yoshio
authors.initials: Y
authors.lastname: Misumi
authors.forename: Stephen J
authors.initials: SJ
authors.lastname: Moss
authors.forename: Masato
authors.initials: M
authors.lastname: Hirata
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D006801:Humans; D007668:Kidney; D051379:Mice; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D052066:N-Ethylmaleimide-Sensitive Proteins; D010446:Peptide Fragments / Q000737:chemistry; D018079:Receptors, GABA / Q000737:chemistry / Q000235:genetics / Q000502:physiology*; D011963:Receptors, GABA-A; D011993:Recombinant Fusion Proteins; D033921:Vesicular Transport Proteins / Q000502:physiology*
chemical_list: C470436:Gabrb2 protein, mouse; D010446:Peptide Fragments; D018079:Receptors, GABA; D011963:Receptors, GABA-A; D011993:Recombinant Fusion Proteins; D033921:Vesicular Transport Proteins; D052066:N-Ethylmaleimide-Sensitive Proteins; C498733:Nsf protein, mouse