Work

GABAB Receptor Constituents Revealed by Tandem Affinity Purification from Transgenic Mice

Year: 2010

Type: article

Abstract: 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.

Source: Journal of Biological Chemistry

DOI: 10.1074/jbc.m109.049700

Authors: Tudor Bartoi, Kristoffer T.G. Rigbolt, Dan Du, Georg Köhr, Blagoy Blagoev, Hans-Christian Kornau

Institutions: Universität Hamburg, University of Southern Denmark, Max Planck Society, Max Planck Institute for Medical Research

Language: English

Topic: Neuroscience and Neuropharmacology Research

Subfield: Cellular and Molecular Neuroscience

Field: Neuroscience

Domain: Life Sciences

Open Access status: hybrid

Estimated APC paid: $3,430

Funder: Deutsche Forschungsgemeinschaft