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Updated: Aug 29, 2025

Methods for the Discovery of Novel Compounds Modulating a Gamma-Aminobutyric Acid Receptor Type A Neurotransmission
Published on: August 16, 2018
GABAA receptor proline 273 at the interdomain interface of the β2 subunit regulates entry into desensitization and
Marek Brodzki1, Jerzy W Mozrzymas2
1Department of Molecular Physiology and Neurobiology, University of Wrocław, ul. Sienkiewicza 21, 50-335 Wrocław, Poland; Department of Biophysics and Neuroscience, Wrocław Medical University, ul. Chałubińskiego 3A, 50-368 Wrocław, Poland.
Aims:
GABAA receptors belong to Cys-loop ion channel family and mediate inhibition in the brain. Despite the abundance of structural data on receptor structure, the molecular scenarios of activation are unknown. In this study we investigated the role of a β2P273 residue in channel gating transitions. This residue is located in a central position of the M2-M3 linker of the interdomain interface, expected to be predisposed to interact with another interfacial element, the β1-β2 loop of the extracellular side. The interactions occurring on this interface have been reported to couple agonist binding to channel gating.
Main Methods:
We recorded micro- and macroscopic current responses of recombinant GABAA receptors mutated at the β2P273 residue (to A, K, E) to saturating GABA. Electrophysiological data served as basis to kinetic modeling, used to decipher which gating transition were affected by mutations.
Key Findings:
Mutations of this residue impaired macroscopic desensitization and accelerated current deactivation with P273E mutant showing greatest deviation from wild-type. Single-channel analysis revealed alterations mainly in short-lived shut times and shortening of openings, resulting in dramatic changes in intraburst open probability. Kinetic modeling indicated that β2P273 mutants show diminished entry into desensitized and open states as well as faster channel closing transitions.
Significance:
In conclusion, we demonstrate that β2P273 of the M2-M3 linker is a crucial element of the ECD-TMD interface regulating the receptor's ability to undergo late gating transitions. Henceforth, this region could be an important target for new pharmacological tools affecting GABAAR-mediated inhibition.
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