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Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels

The concentration of free cytosolic Ca(2+) and the voltage across the plasma membrane are major determinants of cell function. Ca(2+)-permeable non-selective cationic channels are known to regulate these parameters, but understanding of these channels remains inadequate. Here we focus on transient r...

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Autores principales: Rubaiy, Hussein N., Ludlow, Melanie J., Henrot, Matthias, Gaunt, Hannah J., Miteva, Katarina, Cheung, Sin Ying, Tanahashi, Yasuyuki, Hamzah, Nurasyikin, Musialowski, Katie E., Blythe, Nicola M., Appleby, Hollie L., Bailey, Marc A., McKeown, Lynn, Taylor, Roger, Foster, Richard, Waldmann, Herbert, Nussbaumer, Peter, Christmann, Mathias, Bon, Robin S., Muraki, Katsuhiko, Beech, David J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5437225/
https://www.ncbi.nlm.nih.gov/pubmed/28325835
http://dx.doi.org/10.1074/jbc.M116.773556
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author Rubaiy, Hussein N.
Ludlow, Melanie J.
Henrot, Matthias
Gaunt, Hannah J.
Miteva, Katarina
Cheung, Sin Ying
Tanahashi, Yasuyuki
Hamzah, Nurasyikin
Musialowski, Katie E.
Blythe, Nicola M.
Appleby, Hollie L.
Bailey, Marc A.
McKeown, Lynn
Taylor, Roger
Foster, Richard
Waldmann, Herbert
Nussbaumer, Peter
Christmann, Mathias
Bon, Robin S.
Muraki, Katsuhiko
Beech, David J.
author_facet Rubaiy, Hussein N.
Ludlow, Melanie J.
Henrot, Matthias
Gaunt, Hannah J.
Miteva, Katarina
Cheung, Sin Ying
Tanahashi, Yasuyuki
Hamzah, Nurasyikin
Musialowski, Katie E.
Blythe, Nicola M.
Appleby, Hollie L.
Bailey, Marc A.
McKeown, Lynn
Taylor, Roger
Foster, Richard
Waldmann, Herbert
Nussbaumer, Peter
Christmann, Mathias
Bon, Robin S.
Muraki, Katsuhiko
Beech, David J.
author_sort Rubaiy, Hussein N.
collection PubMed
description The concentration of free cytosolic Ca(2+) and the voltage across the plasma membrane are major determinants of cell function. Ca(2+)-permeable non-selective cationic channels are known to regulate these parameters, but understanding of these channels remains inadequate. Here we focus on transient receptor potential canonical 4 and 5 proteins (TRPC4 and TRPC5), which assemble as homomers or heteromerize with TRPC1 to form Ca(2+)-permeable non-selective cationic channels in many mammalian cell types. Multiple roles have been suggested, including in epilepsy, innate fear, pain, and cardiac remodeling, but limitations in tools to probe these channels have restricted progress. A key question is whether we can overcome these limitations and develop tools that are high-quality, reliable, easy to use, and readily accessible for all investigators. Here, through chemical synthesis and studies of native and overexpressed channels by Ca(2+) and patch-clamp assays, we describe compound 31, a remarkable small-molecule inhibitor of TRPC1/4/5 channels. Its potency ranged from 9 to 1300 pm, depending on the TRPC1/4/5 subtype and activation mechanism. Other channel types investigated were unaffected, including TRPC3, TRPC6, TRPV1, TRPV4, TRPA1, TRPM2, TRPM8, and store-operated Ca(2+) entry mediated by Orai1. These findings suggest identification of an important experimental tool compound, which has much higher potency for inhibiting TRPC1/4/5 channels than previously reported agents, impressive specificity, and graded subtype selectivity within the TRPC1/4/5 channel family. The compound should greatly facilitate future studies of these ion channels. We suggest naming this TRPC1/4/5-inhibitory compound Pico145.
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spelling pubmed-54372252017-05-24 Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels Rubaiy, Hussein N. Ludlow, Melanie J. Henrot, Matthias Gaunt, Hannah J. Miteva, Katarina Cheung, Sin Ying Tanahashi, Yasuyuki Hamzah, Nurasyikin Musialowski, Katie E. Blythe, Nicola M. Appleby, Hollie L. Bailey, Marc A. McKeown, Lynn Taylor, Roger Foster, Richard Waldmann, Herbert Nussbaumer, Peter Christmann, Mathias Bon, Robin S. Muraki, Katsuhiko Beech, David J. J Biol Chem Molecular Biophysics The concentration of free cytosolic Ca(2+) and the voltage across the plasma membrane are major determinants of cell function. Ca(2+)-permeable non-selective cationic channels are known to regulate these parameters, but understanding of these channels remains inadequate. Here we focus on transient receptor potential canonical 4 and 5 proteins (TRPC4 and TRPC5), which assemble as homomers or heteromerize with TRPC1 to form Ca(2+)-permeable non-selective cationic channels in many mammalian cell types. Multiple roles have been suggested, including in epilepsy, innate fear, pain, and cardiac remodeling, but limitations in tools to probe these channels have restricted progress. A key question is whether we can overcome these limitations and develop tools that are high-quality, reliable, easy to use, and readily accessible for all investigators. Here, through chemical synthesis and studies of native and overexpressed channels by Ca(2+) and patch-clamp assays, we describe compound 31, a remarkable small-molecule inhibitor of TRPC1/4/5 channels. Its potency ranged from 9 to 1300 pm, depending on the TRPC1/4/5 subtype and activation mechanism. Other channel types investigated were unaffected, including TRPC3, TRPC6, TRPV1, TRPV4, TRPA1, TRPM2, TRPM8, and store-operated Ca(2+) entry mediated by Orai1. These findings suggest identification of an important experimental tool compound, which has much higher potency for inhibiting TRPC1/4/5 channels than previously reported agents, impressive specificity, and graded subtype selectivity within the TRPC1/4/5 channel family. The compound should greatly facilitate future studies of these ion channels. We suggest naming this TRPC1/4/5-inhibitory compound Pico145. American Society for Biochemistry and Molecular Biology 2017-05-19 2017-03-21 /pmc/articles/PMC5437225/ /pubmed/28325835 http://dx.doi.org/10.1074/jbc.M116.773556 Text en © 2017 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/4.0) .
spellingShingle Molecular Biophysics
Rubaiy, Hussein N.
Ludlow, Melanie J.
Henrot, Matthias
Gaunt, Hannah J.
Miteva, Katarina
Cheung, Sin Ying
Tanahashi, Yasuyuki
Hamzah, Nurasyikin
Musialowski, Katie E.
Blythe, Nicola M.
Appleby, Hollie L.
Bailey, Marc A.
McKeown, Lynn
Taylor, Roger
Foster, Richard
Waldmann, Herbert
Nussbaumer, Peter
Christmann, Mathias
Bon, Robin S.
Muraki, Katsuhiko
Beech, David J.
Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title_full Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title_fullStr Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title_full_unstemmed Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title_short Picomolar, selective, and subtype-specific small-molecule inhibition of TRPC1/4/5 channels
title_sort picomolar, selective, and subtype-specific small-molecule inhibition of trpc1/4/5 channels
topic Molecular Biophysics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5437225/
https://www.ncbi.nlm.nih.gov/pubmed/28325835
http://dx.doi.org/10.1074/jbc.M116.773556
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