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

收藏

页码：2563 / 2573

页数：11

相关论文

共 40 条

[31] CFTR, chloride concentration and cell volume: could mammalian protein histidine phosphorylation play a latent role? [J].

Treharne, KJ ;

Crawford, RM ;

Mehta, A .

EXPERIMENTAL PHYSIOLOGY, 2006, 91 (01) :131-139

[32] Identification of mNeonGreen as a pH-Dependent, Turn-On Fluorescent Protein Sensor for Chloride [J].

Tutol, Jasmine N. ;

Kam, Hiu C. ;

Dodani, Sheel C. .

CHEMBIOCHEM, 2019, 20 (14) :1759-1765

[33] Discovery and Characterization of a Naturally Occurring, Turn-On Yellow Fluorescent Protein Sensor for Chloride [J].

Tutol, Jasmine N. ;

Peng, Weicheng ;

Dodani, Sheel C. .

BIOCHEMISTRY, 2019, 58 (01) :31-35

[34] The chloride anion as a signalling effector [J].

Valdivieso, Angel G. ;

Santa-Coloma, Tomas A. .

BIOLOGICAL REVIEWS, 2019, 94 (05) :1839-1856

[35] The Chloride Anion Acts as a Second Messenger in Mammalian Cells - Modifying the Expression of Specific Genes [J].

Valdivieso, Angel G. ;

Clauzure, Mariangeles ;

Massip-Copiz, Macarena ;

Santa-Coloma, Tomas A. .

CELLULAR PHYSIOLOGY AND BIOCHEMISTRY, 2016, 38 (01) :49-64

[36] Sensitivity of the yellow variant of green fluorescent protein to halides and nitrate [J].

Wachter, RM ;

Remington, SJ .

CURRENT BIOLOGY, 1999, 9 (17) :R628-R629

[37] Molecular Mechanism of a Green-Shifted, pH-Dependent Red Fluorescent Protein mKate Variant [J].

Wang, Qi ;

Byrnes, Laura J. ;

Shui, Bo ;

Roehrig, Ute F. ;

Singh, Avtar ;

Chudakov, Dmitriy M. ;

Lukyanov, Sergey ;

Zipfel, Warren R. ;

Kotlikoff, Michael I. ;

Sondermann, Holger .

PLOS ONE, 2011, 6 (08)

[38] mBeRFP, an Improved Large Stokes Shift Red Fluorescent Protein [J].

Yang, Jie ;

Wang, Liang ;

Yang, Fei ;

Luo, Haiming ;

Xu, Lingling ;

Lu, Jinling ;

Zeng, Shaoqun ;

Zhang, Zhihong .

PLOS ONE, 2013, 8 (06)

[39] What biologists want from their chloride reporters - a conversation between chemists and biologists [J].

Zajac, Matthew ;

Chakraborty, Kasturi ;

Saha, Sonali ;

Mahadevan, Vivek ;

Infield, Daniel T. ;

Accardi, Alessio ;

Qiu, Zhaozhu ;

Krishnan, Yamuna .

JOURNAL OF CELL SCIENCE, 2020, 133 (02)

[40] A Genetically-Encoded YFP Sensor with Enhanced Chloride Sensitivity, Photostability and Reduced pH Interference Demonstrates Augmented Transmembrane Chloride Movement by Gerbil Prestin (SLC26a5) [J].

Zhong, Sheng ;

Navaratnam, Dhasakumar ;

Santos-Sacchi, Joseph .

PLOS ONE, 2014, 9 (06)

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