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A Large-Scale High-Throughput Screen for Modulators of SERCA Activity
The sarco/endoplasmic reticulum Ca-ATPase (SERCA) is a P-type ion pump that transports Ca(2+) from the cytosol into the endoplasmic/sarcoplasmic reticulum (ER/SR) in most mammalian cells. It is critically important in muscle, facilitating relaxation and enabling subsequent contraction. Increasing SE...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9776381/ https://www.ncbi.nlm.nih.gov/pubmed/36551215 http://dx.doi.org/10.3390/biom12121789 |
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author | Bidwell, Philip A. Yuen, Samantha L. Li, Ji Berg, Kaja Rebbeck, Robyn T. Aldrich, Courtney C. Roopnarine, Osha Cornea, Razvan L. Thomas, David D. |
author_facet | Bidwell, Philip A. Yuen, Samantha L. Li, Ji Berg, Kaja Rebbeck, Robyn T. Aldrich, Courtney C. Roopnarine, Osha Cornea, Razvan L. Thomas, David D. |
author_sort | Bidwell, Philip A. |
collection | PubMed |
description | The sarco/endoplasmic reticulum Ca-ATPase (SERCA) is a P-type ion pump that transports Ca(2+) from the cytosol into the endoplasmic/sarcoplasmic reticulum (ER/SR) in most mammalian cells. It is critically important in muscle, facilitating relaxation and enabling subsequent contraction. Increasing SERCA expression or specific activity can alleviate muscle dysfunction, most notably in the heart, and we seek to develop small-molecule drug candidates that activate SERCA. Therefore, we adapted an NADH-coupled assay, measuring Ca-dependent ATPase activity of SERCA, to high-throughput screening (HTS) format, and screened a 46,000-compound library of diverse chemical scaffolds. This HTS platform yielded numerous hits that reproducibly alter SERCA Ca-ATPase activity, with few false positives. The top 19 activating hits were further tested for effects on both Ca-ATPase and Ca(2+) transport, in both cardiac and skeletal SR. Nearly all hits increased Ca(2+) uptake in both cardiac and skeletal SR, with some showing isoform specificity. Furthermore, dual analysis of both activities identified compounds with a range of effects on Ca(2+)-uptake and ATPase, which fit into distinct classifications. Further study will be needed to identify which classifications are best suited for therapeutic use. These results reinforce the need for robust secondary assays and criteria for selection of lead compounds, before undergoing HTS on a larger scale. |
format | Online Article Text |
id | pubmed-9776381 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-97763812022-12-23 A Large-Scale High-Throughput Screen for Modulators of SERCA Activity Bidwell, Philip A. Yuen, Samantha L. Li, Ji Berg, Kaja Rebbeck, Robyn T. Aldrich, Courtney C. Roopnarine, Osha Cornea, Razvan L. Thomas, David D. Biomolecules Article The sarco/endoplasmic reticulum Ca-ATPase (SERCA) is a P-type ion pump that transports Ca(2+) from the cytosol into the endoplasmic/sarcoplasmic reticulum (ER/SR) in most mammalian cells. It is critically important in muscle, facilitating relaxation and enabling subsequent contraction. Increasing SERCA expression or specific activity can alleviate muscle dysfunction, most notably in the heart, and we seek to develop small-molecule drug candidates that activate SERCA. Therefore, we adapted an NADH-coupled assay, measuring Ca-dependent ATPase activity of SERCA, to high-throughput screening (HTS) format, and screened a 46,000-compound library of diverse chemical scaffolds. This HTS platform yielded numerous hits that reproducibly alter SERCA Ca-ATPase activity, with few false positives. The top 19 activating hits were further tested for effects on both Ca-ATPase and Ca(2+) transport, in both cardiac and skeletal SR. Nearly all hits increased Ca(2+) uptake in both cardiac and skeletal SR, with some showing isoform specificity. Furthermore, dual analysis of both activities identified compounds with a range of effects on Ca(2+)-uptake and ATPase, which fit into distinct classifications. Further study will be needed to identify which classifications are best suited for therapeutic use. These results reinforce the need for robust secondary assays and criteria for selection of lead compounds, before undergoing HTS on a larger scale. MDPI 2022-11-30 /pmc/articles/PMC9776381/ /pubmed/36551215 http://dx.doi.org/10.3390/biom12121789 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Bidwell, Philip A. Yuen, Samantha L. Li, Ji Berg, Kaja Rebbeck, Robyn T. Aldrich, Courtney C. Roopnarine, Osha Cornea, Razvan L. Thomas, David D. A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title | A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title_full | A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title_fullStr | A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title_full_unstemmed | A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title_short | A Large-Scale High-Throughput Screen for Modulators of SERCA Activity |
title_sort | large-scale high-throughput screen for modulators of serca activity |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9776381/ https://www.ncbi.nlm.nih.gov/pubmed/36551215 http://dx.doi.org/10.3390/biom12121789 |
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