A Superfolder Green Fluorescent Protein-Based Biosensor Allows Monitoring of Chloride in the Endoplasmic Reticulum

被引：11

作者：

Shariati, Kaavian ^{[1
]}

Zhang, Yaohuan ^{[1
,2
]}

Giubbolini, Simone ^{[3
,4
]}

Parra, Riccardo ^{[3
,4
]}

Liang, Steven ^{[1
]}

Edwards, Austin ^{[5
]}

Hejtmancik, J. Fielding ^{[6
]}

Ratto, Gian Michele ^{[3
,4
]}

Arosio, Daniele ^{[7
,8
]}

Ku, Gregory ^{[1
,9
]}

机构：

[1] Univ Calif San Francisco, Diabet Ctr, San Francisco, CA 94143 USA

[2] Univ Calif Berkeley, Dept Nutr Sci & Tox, Metab Biol Grad Program, Berkeley, CA 94720 USA

[3] Consiglio Nazl Ric CNR, Ist Nanosci, Natl Enterprise Nanosci & NanoTechnol NEST, I-56127 Pisa, Italy

[4] Scuola Normale Super Pisa, I-56127 Pisa, Italy

[5] Univ Calif San Francisco, Biol Imaging Dev CoLab, San Francisco, CA 94143 USA

[6] NEI, Ophthalm Genet & Visual Funct Branch, Bethesda, MD 20892 USA

[7] Inst Biophys, CNR, I-38123 Trento, Italy

[8] Univ Trento, CIBIO, I-38123 Trento, Italy

[9] Univ Calif San Francisco, Dept Med, Div Endocrinol, San Francisco, CA 94143 USA

来源：

ACS SENSORS | 2022年 / 7卷 / 08期

关键词：

chloride; endoplasmic reticulum; biosensor; pH; superfolder; INTRACELLULAR CHLORIDE; SECRETORY GRANULES; PH; STRESS; CELLS; GFP;

D O I：

10.1021/acssensors.2c00626

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

Though the concentration of chloride has been measured in the cytoplasm and in secretory granules of live cells, it cannot be measured within the endoplasmic reticulum (ER) due to poor fluorescence of existing biosensors. We developed a fluorescent biosensor composed of a chloride-sensitive superfolder GFP and long Stokes-shifted mKate2 for simultaneous chloride and pH measure-ments that retained fluorescence in the ER lumen. Using this sensor, we showed that the chloride concentration in the ER is significantly lower than that in the cytosol. This improved biosensor enables dynamic measurement of chloride in the ER and may be useful in other environments where protein folding is challenging.

引用

页码：2218 / 2224

页数：7

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