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A Hydrophobic Area of the GABA ρ1 Receptor Containing Phenylalanine 124 Influences Both Receptor Activation and Deactivation

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Abstract

Experimental evidence suggests that GABA ρ1 receptors are potential therapeutic targets for the treatment of a range of neurological conditions, including anxiety and sleep disorders. Homology modelling of the GABA ρ1 extracellular N-terminal domain has revealed a novel hydrophobic area that extends beyond, but not including the GABA-binding site. Phenylalanine 124 (F124) is predicted to be involved in maintaining the structural integrity of the orthosteric-binding site. We have assessed the activity of a series of GABA ρ1 receptors that incorporate a mutation at F124. Wild-type and mutant human GABA ρ1 subunits were expressed in Xenopus laevis oocytes and AD293 cells, and the pharmacology and kinetic properties of the receptors were measured using electrophysiological analysis. Mutation of F124 had minimal effect on receptor pharmacology. However, the rate of deactivation was significantly increased compared to wild type. This study provides further information about the role of residues within a novel hydrophobic area of the GABA ρ1 receptor. This knowledge can help future studies into the design of potent and subtype-selective ligands with therapeutic value.

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Abbreviations

GABA:

γ-Aminobutyric acid

TPMPA:

(1,2,5,6-Tetrahydropyridine-4-yl)methylphosphinic acid

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Acknowledgments

We thank the Xenopus Facility staff for care of frogs. IY acknowledges the support of a John Lamberton Scholarship. JEC acknowledges the support of a UNSW Vice Chancellor’s Research Fellowship.

Conflict of Interest

The authors declare no conflict of interest

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Authors and Affiliations

  1. School of Medical Sciences, UNSW Medicine, The University of New South Wales, Kensington, NSW, 2052, Australia

    J. E. Carland & T. M. Lewis

  2. Current address: Discipline of Pharmacology, School of Medical Sciences, Bosch Institute, Sydney Medical School, The University of Sydney, Sydney, NSW, 2006, Australia

    J. E. Carland

  3. Faculty of Pharmacy, The University of Sydney, Sydney, NSW, 2006, Australia

    I. Yamamoto, J. R. Hanrahan, H. Abdel-Halim, N. Absalom & M. Chebib

  4. Current address: Division of Biophysics and Neurobiology, Department of Molecular Physiology, National Institute for Physiological Sciences, Okazaki, Japan, 444-8585

    I. Yamamoto

Authors
  1. J. E. Carland
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  2. I. Yamamoto
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  3. J. R. Hanrahan
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  4. H. Abdel-Halim
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  5. T. M. Lewis
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  6. N. Absalom
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  7. M. Chebib
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Corresponding author

Correspondence to M. Chebib.

Additional information

Jane E. Carland and Izumi Yamamoto made equal contributions to this work.

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Carland, J.E., Yamamoto, I., Hanrahan, J.R. et al. A Hydrophobic Area of the GABA ρ1 Receptor Containing Phenylalanine 124 Influences Both Receptor Activation and Deactivation. J Mol Neurosci 55, 305–313 (2015). https://doi.org/10.1007/s12031-014-0322-7

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  • DOI: https://doi.org/10.1007/s12031-014-0322-7

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