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Title: Inactivation-mimicking block of the epithelial calcium channel TRPV6

Abstract

Epithelial calcium channel TRPV6 plays vital roles in calcium homeostasis, and its dysregulation is implicated in multifactorial diseases, including cancers. Here, we study the molecular mechanism of selective nanomolar-affinity TRPV6 inhibition by (4-phenylcyclohexyl)piperazine derivatives (PCHPDs). We use x-ray crystallography and cryo–electron microscopy to solve the inhibitor-bound structures of TRPV6 and identify two types of inhibitor binding sites in the transmembrane region: (i) modulatory sites between the S1-S4 and pore domains normally occupied by lipids and (ii) the main site in the ion channel pore. Our structural data combined with mutagenesis, functional and computational approaches suggest that PCHPDs plug the open pore of TRPV6 and convert the channel into a nonconducting state, mimicking the action of calmodulin, which causes inactivation of TRPV6 channels under physiological conditions. This mechanism of inhibition explains the high selectivity and potency of PCHPDs and opens up unexplored avenues for the design of future-generation biomimetic drugs.

Authors:
ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3];  [3];  [4]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [1]; ORCiD logo [2]; ORCiD logo [3]
  1. Univ. of Bern (Switzerland)
  2. Johannes Kepler Univ. Linz (Austria)
  3. Columbia Univ., New York, NY (United States)
  4. Columbia Univ., New York, NY (United States); Indian Inst. of Technology (IIT), Kanpur (India)
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE
OSTI Identifier:
1729717
Resource Type:
Accepted Manuscript
Journal Name:
Science Advances
Additional Journal Information:
Journal Volume: 6; Journal Issue: 48; Journal ID: ISSN 2375-2548
Publisher:
AAAS
Country of Publication:
United States
Language:
ENGLISH
Subject:
59 BASIC BIOLOGICAL SCIENCES

Citation Formats

Bhardwaj, Rajesh, Lindinger, Sonja, Neuberger, Arthur, Nadezhdin, Kirill D., Singh, Appu K., Cunha, Micael R., Derler, Isabella, Gyimesi, Gergely, Reymond, Jean-Louis, Hediger, Matthias A., Romanin, Christoph, and Sobolevsky, Alexander I.. Inactivation-mimicking block of the epithelial calcium channel TRPV6. United States: N. p., 2020. Web. https://doi.org/10.1126/sciadv.abe1508.
Bhardwaj, Rajesh, Lindinger, Sonja, Neuberger, Arthur, Nadezhdin, Kirill D., Singh, Appu K., Cunha, Micael R., Derler, Isabella, Gyimesi, Gergely, Reymond, Jean-Louis, Hediger, Matthias A., Romanin, Christoph, & Sobolevsky, Alexander I.. Inactivation-mimicking block of the epithelial calcium channel TRPV6. United States. https://doi.org/10.1126/sciadv.abe1508
Bhardwaj, Rajesh, Lindinger, Sonja, Neuberger, Arthur, Nadezhdin, Kirill D., Singh, Appu K., Cunha, Micael R., Derler, Isabella, Gyimesi, Gergely, Reymond, Jean-Louis, Hediger, Matthias A., Romanin, Christoph, and Sobolevsky, Alexander I.. Fri . "Inactivation-mimicking block of the epithelial calcium channel TRPV6". United States. https://doi.org/10.1126/sciadv.abe1508. https://www.osti.gov/servlets/purl/1729717.
@article{osti_1729717,
title = {Inactivation-mimicking block of the epithelial calcium channel TRPV6},
author = {Bhardwaj, Rajesh and Lindinger, Sonja and Neuberger, Arthur and Nadezhdin, Kirill D. and Singh, Appu K. and Cunha, Micael R. and Derler, Isabella and Gyimesi, Gergely and Reymond, Jean-Louis and Hediger, Matthias A. and Romanin, Christoph and Sobolevsky, Alexander I.},
abstractNote = {Epithelial calcium channel TRPV6 plays vital roles in calcium homeostasis, and its dysregulation is implicated in multifactorial diseases, including cancers. Here, we study the molecular mechanism of selective nanomolar-affinity TRPV6 inhibition by (4-phenylcyclohexyl)piperazine derivatives (PCHPDs). We use x-ray crystallography and cryo–electron microscopy to solve the inhibitor-bound structures of TRPV6 and identify two types of inhibitor binding sites in the transmembrane region: (i) modulatory sites between the S1-S4 and pore domains normally occupied by lipids and (ii) the main site in the ion channel pore. Our structural data combined with mutagenesis, functional and computational approaches suggest that PCHPDs plug the open pore of TRPV6 and convert the channel into a nonconducting state, mimicking the action of calmodulin, which causes inactivation of TRPV6 channels under physiological conditions. This mechanism of inhibition explains the high selectivity and potency of PCHPDs and opens up unexplored avenues for the design of future-generation biomimetic drugs.},
doi = {10.1126/sciadv.abe1508},
journal = {Science Advances},
number = 48,
volume = 6,
place = {United States},
year = {2020},
month = {11}
}

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