New Red-Emitting Chloride-Sensitive Fluorescent Protein with Biological Uses
Metadatos
Afficher la notice complèteAuteur
Salto González, Rafael; Girón González, María Dolores; Puente Muñoz, Virginia; Vílchez Rienda, José Dámaso; Espinar Barranco, Laura; Valverde Pozo, Javier; Paredes Martínez, José ManuelEditorial
American Chemical Society
Materia
Red fluorescent proteins Chloride sensors Imaging Two-photon excitation microscopy Genetically encoded sensors Intracellular sensors
Date
2021-06-21Referencia bibliográfica
ACS Sens. 2021, 6, 2563−2573. [https://doi.org/10.1021/acssensors.1c00094]
Patrocinador
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 2018.256Résumé
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.