期刊
ACS SENSORS
卷 -, 期 -, 页码 -出版社
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.2c00626
关键词
chloride; endoplasmic reticulum; biosensor; pH; superfolder
资金
- NIH [R01 DK118337-01, R01 DK107650-01, R03 DK124904-01, P30 DK063720]
Researchers have developed an improved fluorescent biosensor that can simultaneously measure chloride ions and pH in the endoplasmic reticulum (ER), which is useful for detecting chloride concentration in environments with challenging protein folding.
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.
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