GABAB Receptor Constituents Revealed by Tandem Affinity Purification from Transgenic Mice

被引:38
作者
Bartoi, Tudor
Rigbolt, Kristoffer T. G. [1 ]
Du, Dan [2 ]
Koehr, Georg [2 ]
Blagoev, Blagoy [1 ]
Kornau, Hans-Christian
机构
[1] Univ So Denmark, Dept Biochem & Mol Biol, CEBI, DK-5230 Odense M, Denmark
[2] Max Planck Inst Med Res, Dept Mol Neurobiol, D-69120 Heidelberg, Germany
关键词
SUBCELLULAR-LOCALIZATION; STRUCTURAL-ANALYSIS; STRESS INDUCTION; MESSENGER-RNA; T1; DOMAIN; PROTEINS; CLONING; GENE; EXPRESSION; RAT;
D O I
10.1074/jbc.M109.049700
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:20625 / 20633
页数:9
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