Single-channel electrophysiology with lipid bilayer systems requires ion channel expression, purification from cell culture, and reconstitution in proteoliposomes for delivery to a planar bilayer. Here we demonstrate that single-channel current measurements of the potassium channels KcsA and hERGS5-S6 can be obtained by direct insertion in interdroplet lipid bilayers from microliters of a cell-free expression medium.
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Methods
January 2025
Institute of Cardiology, Lithuanian University of Health Sciences, Kaunas 50103, Lithuania; Department of Mathematical Modelling, Kaunas University of Technology, Kaunas 51368, Lithuania.
Gap junction (GJ) channels, formed of connexin (Cx) protein, enable direct intercellular communication in most vertebrate tissues. One of the key biophysical characteristics of these channels is their unitary conductance, which can be affected by mutations in Cx genes and various biochemical factors, such as posttranslational modifications. Due to the unique intercellular configuration of GJ channels, recording single-channel currents is challenging, and precise data on unitary conductances of some Cx isoforms remain limited.
View Article and Find Full Text PDFBiophys J
December 2024
Department of Pharmacology, Larner College of Medicine, Vermont Center for Cardiovascular and Brain Health, University of Vermont, Burlington, Vermont. Electronic address:
Amyloid beta (Aβ) peptide accumulation on blood vessels in the brain is a hallmark of neurodegeneration. While Aβ peptides constrict cerebral arteries and arterioles, their impact on capillaries is less understood. Aβ was recently shown to constrict brain capillaries through pericyte contraction, but whether-and if so how-Aβ affects endothelial cells (ECs) remains unknown.
View Article and Find Full Text PDFJ Phys Chem B
December 2024
Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, Oxford, Mississippi 38677, United States.
bioRxiv
August 2024
Department of Biomolecular Sciences, School of Pharmacy, University of Mississippi, Oxford, MS.
Polyunsaturated fatty acids (PUFAs) and their analogs play a significant role in modulating the activity of diverse ion channels, and recent studies show that these lipids potentiate acid-sensing ion channels (ASICs), leading to increased activity. The potentiation of the channel stems from multiple gating changes, but the exact mechanism of these effects remains uncertain. We posit a mechanistic explanation for one of these changes in channel function, the increase in the maximal current, by applying a combination of electrophysiology and all-atom molecular dynamics simulations on the open-state hASIC3.
View Article and Find Full Text PDFJ Biol Chem
October 2024
Department of Dermatology, Hangzhou Third People's Hospital, Affiliated Hangzhou Dermatology Hospital of Zhejiang University School of Medicine, Hangzhou, China. Electronic address:
Transient receptor potential vanilloid 3 channel (TRPV3) is closely associated with skin inflammation, but there is a lack of effective and specific inhibitors for clinical use. In this study, we identified antimalarial hydroxychloroquine (HCQ) as a selective TRPV3 inhibitor following the prediction by network pharmacology data analysis. In whole-cell patch-clamp recordings, HCQ inhibited the current of the TRPV3 channel, with an IC50 of 51.
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