Coupling of GABAB receptor GABAB2 subunit to G proteins: Evidence from Xenopus oocyte and baby hamster kidney cell expression system

Yasuhito Uezono, Masato Kanaide, Muneshige Kaibara, Rachel Barzilai, Nathan Dascal, Koji Sumikawa, Kohtaro Taniyama

Research output: Contribution to journalArticlepeer-review

Abstract

Coupling of functional GABAB receptors (GABABR) to G proteins was investigated with an expression system of baby hamster kidney (BHK) cells and Xenopus oocytes. Fluorescence resonance energy transfer (FRET) analysis of BHK cells coexpressing GABAB1a receptor (GB 1aR) fused to Cerulean, a brighter variant of cyan fluorescent protein, and GABAB2 receptor (GB2R) fused to Venus, a brighter variant of yellow fluorescent protein, revealed that GB 1aR-Cerulean and GB2R-Venus form a heterodimer. The GABABR agonists baclofen and 3-aminopropylphosphonic acid (3-APPA) elicited inward-rectifying K+ currents in a concentration-dependent manner in oocytes expressing GB1aR and GB2R, or GB 1aR-Cerulean and GB2R-Venus, together with G protein-activated inward-rectifying K+ channels (GIRKs), but not in oocytes expressing GB1aR alone or GB2R alone together with GIRKs. Oocytes coexpressing GB1aR + Gαi2- fused GB2R (GB2R-Gαi2) caused faster K + currents in response to baclofen. Furthermore, oocytes coexpressing GB1aR + GB2R fused to Gαqi5 (a chimeric Gαq protein that activates PLC pathways) caused PLC-mediated Ca2+-activated Cl- currents in response to baclofen. In contrast, these responses to baclofen were not observed in oocytes coexpressing GB1aR-Gαi2 or GB1aR- Gαqi5 together with GB2R. BHK cells and Xenopus oocytes coexpressing GB1aR-Cerulean + a triplet tandem of GB 2R-Venus-Gαqi5 caused FRET and Ca 2+-activated Cl- currents, respectively, with a similar potency in BHK cells coexpressing GB1aR-Cerulean + GB 2R-Venus and in oocytes coexpressing GB1aR + GB 2R-Gαqi5. Our results indicate that functional GABABR forms a heterodimer composed of GB1R and GB 2R and that the signal transducing G proteins are directly coupled to GB2R but not to GB1R.

Original languageEnglish
Pages (from-to)C200-C207
JournalAmerican Journal of Physiology - Cell Physiology
Volume290
Issue number1
DOIs
StatePublished - Jan 2006

Keywords

  • Fluorescence resonance energy transfer

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