A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum

[Image: see text] Though the concentration of chloride has been measured in the cytoplasm and in secretory granules of live cells, it cannot be measured within the endoplasmic reticulum (ER) due to poor fluorescence of existing biosensors. We developed a fluorescent biosensor composed of a chloride-...

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Autores principales: Shariati, Kaavian, Zhang, Yaohuan, Giubbolini, Simone, Parra, Riccardo, Liang, Steven, Edwards, Austin, Hejtmancik, J. Fielding, Ratto, Gian Michele, Arosio, Daniele, Ku, Gregory
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2022
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9425558/
https://www.ncbi.nlm.nih.gov/pubmed/35951356
http://dx.doi.org/10.1021/acssensors.2c00626
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author Shariati, Kaavian
Zhang, Yaohuan
Giubbolini, Simone
Parra, Riccardo
Liang, Steven
Edwards, Austin
Hejtmancik, J. Fielding
Ratto, Gian Michele
Arosio, Daniele
Ku, Gregory
author_facet Shariati, Kaavian
Zhang, Yaohuan
Giubbolini, Simone
Parra, Riccardo
Liang, Steven
Edwards, Austin
Hejtmancik, J. Fielding
Ratto, Gian Michele
Arosio, Daniele
Ku, Gregory
author_sort Shariati, Kaavian
collection PubMed
description [Image: see text] Though the concentration of chloride has been measured in the cytoplasm and in secretory granules of live cells, it cannot be measured within the endoplasmic reticulum (ER) due to poor fluorescence of existing biosensors. We developed a fluorescent biosensor composed of a chloride-sensitive superfolder GFP and long Stokes-shifted mKate2 for simultaneous chloride and pH measurements that retained fluorescence in the ER lumen. Using this sensor, we showed that the chloride concentration in the ER is significantly lower than that in the cytosol. This improved biosensor enables dynamic measurement of chloride in the ER and may be useful in other environments where protein folding is challenging.
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spelling pubmed-94255582022-08-31 A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum Shariati, Kaavian Zhang, Yaohuan Giubbolini, Simone Parra, Riccardo Liang, Steven Edwards, Austin Hejtmancik, J. Fielding Ratto, Gian Michele Arosio, Daniele Ku, Gregory ACS Sens [Image: see text] Though the concentration of chloride has been measured in the cytoplasm and in secretory granules of live cells, it cannot be measured within the endoplasmic reticulum (ER) due to poor fluorescence of existing biosensors. We developed a fluorescent biosensor composed of a chloride-sensitive superfolder GFP and long Stokes-shifted mKate2 for simultaneous chloride and pH measurements that retained fluorescence in the ER lumen. Using this sensor, we showed that the chloride concentration in the ER is significantly lower than that in the cytosol. This improved biosensor enables dynamic measurement of chloride in the ER and may be useful in other environments where protein folding is challenging. American Chemical Society 2022-08-11 2022-08-26 /pmc/articles/PMC9425558/ /pubmed/35951356 http://dx.doi.org/10.1021/acssensors.2c00626 Text en © 2022 The Authors. Published by American Chemical Society https://creativecommons.org/licenses/by/4.0/Permits the broadest form of re-use including for commercial purposes, provided that author attribution and integrity are maintained (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Shariati, Kaavian
Zhang, Yaohuan
Giubbolini, Simone
Parra, Riccardo
Liang, Steven
Edwards, Austin
Hejtmancik, J. Fielding
Ratto, Gian Michele
Arosio, Daniele
Ku, Gregory
A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title_full A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title_fullStr A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title_full_unstemmed A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title_short A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum
title_sort superfolder green fluorescent protein-based biosensor allows monitoring of chloride in the endoplasmic reticulum
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9425558/
https://www.ncbi.nlm.nih.gov/pubmed/35951356
http://dx.doi.org/10.1021/acssensors.2c00626
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