# A single main-chain hydrogen bond required to keep GABAA receptors closed

## TL;DR

The study identifies a single hydrogen bond in GABAA receptors that keeps them closed until neurotransmitter activation.

## Contribution

The discovery of a specific main-chain hydrogen bond in the β2 subunit critical for channel closure and opening.

## Key findings

- A single main-chain H-bond in the β2 subunit M2-M3 linker is required to keep the channel closed.
- Breaking this H-bond accounts for about one third of the energy needed to open the channel during GABA activation.
- The analogous H-bond in the α1 subunit does not affect channel gating.

## Abstract

GABAA receptors (GABAARs) are the primary inhibitory neurotransmitter receptors throughout the central nervous system. Genetic mutations causing their dysfunction are related to a broad spectrum of human disorders such as epilepsy, neurodevelopment and intellectual disability, autism spectrum disorder, schizophrenia, and depression. GABAARs are also important drug targets for anxiolytics, anticonvulsants, antidepressants, and anesthetics. Despite significant progress in understanding their three-dimensional structure, a critical gap remains in determining the molecular basis for channel gating. We recently identified mutations in the M2-M3 linkers that suggest linker flexibility has asymmetric subunit-specific correlations with channel opening. Here we use non-canonical amino acids (ncAAs) to investigate the role of main-chain H-hydrogen bonds (H-bonds) that may stabilize the M2-M3 linkers. We show that a single main-chain H-bond within the β2 subunit M2-M3 linker inhibits pore opening and is required to keep the unliganded channel closed. Furthermore, breaking this H-bond accounts for approximately one third of the energy used to open the channel during activation by GABA. In contrast, the analogous H-bond in the α1 subunit has no effect on gating. Our molecular simulations support the idea that channel opening involves the state-dependent breakage/disruption of a specific main-chain H-bond within the β2 subunit M2-M3 linker.

The authors identify a single main-chain hydrogen bond required to keep GABAA receptors closed in the absence of neurotransmitter. Electrophysiology and molecular dynamics simulations suggest disruption of this bond is a key component of channel opening during inhibitory synaptic signaling in the brain.