New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
| Autor: | Jose M. Paredes, Laura Espinar-Barranco, Virginia Puente-Muñoz, Jose D. Vílchez, María D. Girón, Javier Valverde-Pozo, Daniele Arosio, Rafael Salto |
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| Rok vydání: | 2021 |
| Předmět: |
Intracellular sensors
Bioengineering chloride sensors 02 engineering and technology Red fluorescent proteins genetically encoded sensors 01 natural sciences Chloride Genetically encoded sensors Article Green fluorescent protein Imaging Chlorides medicine intracellular sensors Fluorescent protein red fluorescent proteins Instrumentation Two-photon excitation microscopy Fluid Flow and Transfer Processes imaging two-photon excitation microscopy Chemistry Process Chemistry and Technology Dual emission 010401 analytical chemistry Mutagenesis Chromophore Hydrogen-Ion Concentration 021001 nanoscience & nanotechnology Chloride sensors 0104 chemical sciences Luminescent Proteins Biophysics Mutagenesis Site-Directed 0210 nano-technology Isomerization Intracellular medicine.drug |
| Zdroj: | ACS Sensors ACS sensors Online 6 (2021): 2563–2573. doi:10.1021/acssensors.1c00094 info:cnr-pdr/source/autori:Salto R.; Giron M.D.; Puente-Munoz V.; Vilchez J.D.; Espinar-Barranco L.; Valverde-Pozo J.; Arosio D.; Paredes J.M./titolo:New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses/doi:10.1021%2Facssensors.1c00094/rivista:ACS sensors Online/anno:2021/pagina_da:2563/pagina_a:2573/intervallo_pagine:2563–2573/volume:6 Digibug. Repositorio Institucional de la Universidad de Granada instname |
| ISSN: | 2379-3694 |
| Popis: | This work was financially supported by CTQ2017-85685-R, CTQ2017-85454-C2-1-P, and CTQ2017-86125-P (MICIU/AEI/ERDF). J.M.P. and J.D.V. thank UEQ for funding. J.V.P. is supported by an FPU fellowship (FPU17/04749). D.A. was supported by the Italian Cystic Fibrosis Foundation (grant FFC#3/2019) and by Fondazione Cassa Rurale Trento Rovereto (ref 2018.256). The authors acknowledge the Universidad of Granada (Spain) cell culture and microscopy central facilities (CIC-UGR). A new chloride-sensitive red fluorescent protein derived from Entacmaea quadricolor is described. We found that mBeRFP exhibited moderate sensitivity to chloride and, via sitedirected mutagenesis (S94V and R205Y), we increased the chloride affinity by more than an order of magnitude (kd = 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. MICIU/AEI/ERDF CTQ2017-85685-R CTQ2017-85454-C2-1-P CTQ2017-86125-P UEQ Spanish Government FPU17/04749 Ministry of Health, Italy Italian Cystic Fibrosis Research Foundation 3/2019 Fondazione Cassa Rurale Trento Rovereto 2018.256 |
| Databáze: | OpenAIRE |
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