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

被引：8

作者：

Salto, Rafael ^{[1
]}

Giron, Maria D. ^{[1
]}

Puente-Munoz, Virginia ^{[2
]}

Vilchez, Jose D. ^{[1
]}

Espinar-Barranco, Laura ^{[2
]}

Valverde-Pozo, Javier ^{[2
]}

Arosio, Daniele ^{[3
]}

Paredes, Jose M. ^{[2
]}

机构：

[1] Univ Granada, Unidad Excelencia Quim Aplicada Biomed & Medioamb, Fac Pharm, Dept Biochem & Mol Biol 2, Granada 18071, Spain

[2] Univ Granada, Unidad Excelencia Quim Aplicada Biomed & Medioamb, Fac Pharm, Dept Phys Chem, Granada 18071, Spain

[3] CNR, Ist Biofis IBF, I-38123 Trento, Italy

来源：

ACS SENSORS | 2021年 / 6卷 / 07期

关键词：

red fluorescent proteins; chloride sensors; imaging; two-photon excitation microscopy; genetically encoded sensors; intracellular sensors; INTRACELLULAR CHLORIDE; CYSTIC-FIBROSIS; PH; ISOMERIZATION; INDICATOR; VARIANT; CELLS;

D O I：

10.1021/acssensors.1c00094

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

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 (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.

引用

页码：2563 / 2573

页数：11

共 40 条

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