Direct measurements of neurosteroid binding to specific sites on GABA_A receptors.

ABSTRACT:

Background and purpose

Neurosteroids are allosteric modulators of GABA_A currents, acting through several functional binding sites although their affinity and specificity for each site are unknown. The goal of this study was to measure steady-state binding affinities of various neurosteroids for specific sites on the GABA_A receptor.

Experimental approach

Two methods were developed to measure neurosteroid binding affinity: (1) quenching of specific tryptophan residues in neurosteroid binding sites by the neurosteroid 17-methylketone group, and (2) FRET between MQ290 (an intrinsically fluorescent neurosteroid) and tryptophan residues in the binding sites. The assays were developed using ELIC-α1GABA_AR, a chimeric receptor containing transmembrane

SUBMITTER: Chintala SM 

PROVIDER: S-EPMC11585924 | biostudies-literature | 2024 Nov

REPOSITORIES: biostudies-literature