Biology subjects

Brams, M.

Publications and source records attributed to Brams, M..

4 recordsLinked to original sources

bioRxiv2025

Structural basis for gating inhibition by the cytoplasmic domain in HCN1 channels

Hyperpolarization-activated, cyclic nucleotide-gated (HCN) channels generate rhythmic electrical activity in cardiac and neuronal tissues, with isoform-specific cAMP sensitivity remaining poorly understood. While HCN2 exhibits strong cAMP regulation, HCN1 shows minimal response. To investigate the structural basis of this divergence, we analyzed two engineered HCN1 variants using cryo-electron microscopy. One variant (HCN112) incorporates the C-linker and CNBD from HCN2 into the HCN1 backbone and exhibited enhanced cAMP sensitivity, with structural analysis revealing a compressed cytoplasmic domain arrangement that may facilitate regulatory interactions. In contrast, the truncated HCN1{Delta}C variant (lacking the cytoplasmic domain) displayed an intermediately open pore conformation, supporting auto-inhibitory regulation by the CNBD in wild-type channels. These structural insights elucidate how domain-specific interactions modulate cAMP-dependent gating and intrinsic auto-inhibition, resolving long-standing questions about mechanistic divergence among HCN isoforms. Our findings not only shed new light on the structural mechanisms underlying isoform-specific cAMP sensitivity but also have implications for the development of therapeutic strategies targeting HCN channels in neurological and cardiac disorders. SIGNIFICANCE STATEMENTHyperpolarization-activated cyclic nucleotide-gated (HCN) channels are essential regulators of rhythmic electrical activity in the heart and brain, and their dysfunction is linked to disorders such as epilepsy, depression, and cardiac arrhythmias. Although HCN channel isoforms display highly divergent responses to cAMP modulation, the structural basis for these differences has remained elusive. This study reveals, through cryo-EM structures, how domain-specific interactions within the cytoplasmic regions of HCN1 channels underlie their unique gating and auto-inhibition properties, as well as their muted cAMP sensitivity compared to HCN2. These insights resolve long-standing mechanistic questions about HCN channel regulation and pave the way for rational design of targeted therapies that selectively modulate HCN isoform activity in neurological and cardiac disease.

biochemistry↗

bioRxiv2025

Structure of a pH-sensitive pentameric ligand-gated ion channel from the Sarcoptes scabies mite

Scabies is a skin infestation caused by the Sarcoptes scabiei mite. It produces a substantial global health burden, which is exacerbated by emerging resistance to prominent treatments, such as ivermectin. An anionic pentameric ligand-gated ion channel (pLGIC) identified in the mite, termed as SsCl, shows unique pH-sensitivity and is significantly modulated by ivermectin. Here, we use cryo-EM and electrophysiology to explore the pH-sensing mechanisms of SsCl and the impact of ivermectin on channel activity. Structures of SsCl were resolved under various pH conditions to capture a closed (pH 6.5) and desensitized (pH 9) state, alongside ivermectin-bound conformations. The desensitized structure adopts an unexpected "hourglass" conformation, suggesting the gating mechanisms may be more related to cation-selective channels. Structure-based analysis and mutagenesis highlighted extracellular histidine and glutamate residues which act as pH-sensors, revealing the protonation-dependent gating mechanism. Ivermectin binds at transmembrane subunit interfaces, stabilizing an open-pore conformation through hydrophobic interactions. Ivermectin bound-structures reveal a pH-dependent modulation, enhancing open-state prevalence at pH 9 and enabling atypical activation at pH 6.5, consistent with electrophysiological data. These structural and functional insights elucidate SsCls unique pH sensitivity and ivermectins mode of action, providing a foundation for designing next-generation therapeutics targeting this pathogen-related ion channel.

biochemistry↗

bioRxiv2021

A cationic lipid site at the outward transmembrane face of a pentameric ligand-gated ion channel

Pentameric ligand-gated ion channels (pLGICs) are crucial mediators of electrochemical signal transduction from bacteria to humans. Lipids play an important role in regulating pLGIC function, yet the structural basis for specific pLGIC-lipid interactions remains poorly understood. The bacterial channel ELIC recapitulates several properties of eukaryotic pLGICs, including activation by the neurotransmitter GABA and sensitivity to lipids, offering a simplified model system for structure-function studies. In this study, functional effects of non-canonical amino acid substitution of W206 at the top of the M1-helix, combined with detergent interactions observed in recent X-ray structures, are consistent with this region being the location of a lipid binding site on the outward face of the ELIC transmembrane domain. Coarse-grained and atomistic molecular dynamics simulations revealed preferential binding of lipids containing a positive charge, particularly involving interactions with residue W206 consistent with cation-{pi} binding. Polar contacts from the principal subunit, particularly M3 residue Q264, further supported lipid binding via headgroup ester linkages. Aromatic residues were identified at analogous sites in a handful of eukaryotic family members, including the human GABAA receptor subunit {varepsilon}, suggesting conservation of relevant interactions in other evolutionary branches. Further mutagenesis experiments indicated that mutations at this site in {varepsilon}-containing GABAA receptors can change the apparent affinity of the agonist response to GABA, consistent with a potential role of this site in channel gating. In conclusion, this work is a detailed case study in type-specific lipid interactions at an evolutionarily distinctive pLGIC site, with implications for lipid modulation and lipophilic drug design.

biophysics↗

bioRxiv2019

A functionally conserved mechanism of modulation via a vestibule site in pentameric ligand-gated ion channels

Pentameric ligand-gated ion channels (pLGICs) belong to a class of ion channels involved in fast synaptic signaling in the central and peripheral nervous systems. Molecules acting as allosteric modulators target binding sites that are remote from the neurotransmitter binding site, but functionally affect coupling of ligand binding to channel opening. Here, we investigated an allosteric binding site in the ion channel vestibule, which has converged from a series of studies on prokaryote and eukaryote channel homologs. We discovered single domain antibodies, called nanobodies, which are functionally active as allosteric modulators, and solved co-crystal structures of the prokaryote channel ELIC bound either to a positive (PAM) or a negative (NAM) allosteric modulator. We extrapolate the functional importance of the vestibule binding site to eukaryote ion channels, suggesting a conserved mechanism of allosteric modulation. This work identifies key elements of allosteric binding sites and extends drug design possibilities in pLGICs using nanobodies.

neuroscience↗