GABAB receptor constituents revealed by tandem affinity purification from transgenic mice.
GABA(B) receptors function as heterodimeric G-protein-coupled receptors for the neurotransmitter gamma-aminobutyric acid (GABA). Receptor subtypes, based on isoforms of the ligand-binding subunit GABA(B1), are thought to involve a differential set of associated proteins. Here, we describe two mouse lines that allow a straightforward biochemical isolation of GABA(B) receptors. The transgenic mice express GABA(B1) isoforms that contain sequences for a two-step affinity purification, in addition to their endogenous subunit repertoire. Comparative analyses of purified samples from the transgenic mice and wild-type control animals revealed two novel components of the GABA(B1) complex. One of the identified proteins, potassium channel tetramerization domain-containing protein 12, associates with heterodimeric GABA(B) receptors via the GABA(B2) subunit. In transfected hippocampal neurons, potassium channel tetramerization domain-containing protein 12 augmented axonal surface targeting of GABA(B2). The mice equipped with tags on GABA(B1) facilitate validation and identification of native binding partners of GABA(B) receptors, providing insight into the molecular mechanisms of synaptic modulation.
Bartoi
Tudor
T
Center for Molecular Neurobiology (ZMNH), University of Hamburg, Falkenried 94, D-20251 Hamburg, Germany.
Rigbolt
Kristoffer T G
KT
Du
Dan
D
Köhr
Georg
G
Blagoev
Blagoy
B
Kornau
Hans-Christian
HC
D000331:Aequorin / Q000235:genetics; D000818:Animals; D015153:Blotting, Western; D001921:Brain / Q000502:physiology; D016466:CHO Cells; D022202:Chromosomes, Artificial, Bacterial / Q000235:genetics; D003001:Cloning, Molecular; D006224:Cricetinae; D003412:Cricetulus; D005091:Exons / Q000235:genetics; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins / Q000235:genetics; D017403:In Situ Hybridization; D051379:Mice; D008822:Mice, Transgenic; D009474:Neurons / Q000502:physiology; D051381:Rats; D018080:Receptors, GABA-B / Q000235:genetics / Q000502:physiology*; D014162:Transfection
D018080:Receptors, GABA-B; D049452:Green Fluorescent Proteins; D000331:Aequorin
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