New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses

[Image: see text] A new chloride-sensitive red fluorescent protein derived from Entacmaea quadricolor is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via site-directed mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitu...

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Autores principales: Salto, Rafael, Giron, Maria D., Puente-Muñoz, Virginia, Vilchez, Jose D., Espinar-Barranco, Laura, Valverde-Pozo, Javier, Arosio, Daniele, Paredes, Jose M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Chemical Society 2021
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8478333/
https://www.ncbi.nlm.nih.gov/pubmed/34148347
http://dx.doi.org/10.1021/acssensors.1c00094
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author Salto, Rafael
Giron, Maria D.
Puente-Muñoz, Virginia
Vilchez, Jose D.
Espinar-Barranco, Laura
Valverde-Pozo, Javier
Arosio, Daniele
Paredes, Jose M.
author_facet Salto, Rafael
Giron, Maria D.
Puente-Muñoz, Virginia
Vilchez, Jose D.
Espinar-Barranco, Laura
Valverde-Pozo, Javier
Arosio, Daniele
Paredes, Jose M.
author_sort Salto, Rafael
collection PubMed
description [Image: see text] A new chloride-sensitive red fluorescent protein derived from Entacmaea quadricolor is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via site-directed mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitude (k(d) = 106 ± 6 mM) at physiological pH. In addition, cis–trans isomerization of the chromophore produces a dual emission band with different chloride sensitivities, which allowed us to develop a ratiometric methodology to measure intracellular chloride concentrations.
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spelling pubmed-84783332021-09-29 New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses Salto, Rafael Giron, Maria D. Puente-Muñoz, Virginia Vilchez, Jose D. Espinar-Barranco, Laura Valverde-Pozo, Javier Arosio, Daniele Paredes, Jose M. ACS Sens [Image: see text] A new chloride-sensitive red fluorescent protein derived from Entacmaea quadricolor is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via site-directed mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitude (k(d) = 106 ± 6 mM) at physiological pH. In addition, cis–trans isomerization of the chromophore produces a dual emission band with different chloride sensitivities, which allowed us to develop a ratiometric methodology to measure intracellular chloride concentrations. American Chemical Society 2021-06-21 2021-07-23 /pmc/articles/PMC8478333/ /pubmed/34148347 http://dx.doi.org/10.1021/acssensors.1c00094 Text en © 2021 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 Salto, Rafael
Giron, Maria D.
Puente-Muñoz, Virginia
Vilchez, Jose D.
Espinar-Barranco, Laura
Valverde-Pozo, Javier
Arosio, Daniele
Paredes, Jose M.
New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title_full New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title_fullStr New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title_full_unstemmed New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title_short New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
title_sort new red-emitting chloride-sensitive fluorescent protein with biological uses
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8478333/
https://www.ncbi.nlm.nih.gov/pubmed/34148347
http://dx.doi.org/10.1021/acssensors.1c00094
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