期刊
BIOCHEMISTRY
卷 58, 期 1, 页码 31-35出版社
AMER CHEMICAL SOC
DOI: 10.1021/acs.biochem.8b00928
关键词
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资金
- University of Texas at Dallas
- Welch Foundation [AT -1918-20170325]
- National Institutes of Health [1R3SGM128923-01]
Fluorescent proteins have been extensively engineered and applied as optical indicators for chloride in a variety of biological contexts. Surprisingly, given the biodiversity of fluorescent proteins, a naturally occurring chloride sensor has not been reported to date. Here, we present the identification and spectroscopic characterization of the yellow fluorescent protein from the jellyfish Phialidium sp. (phiYFP), a rare example of a naturally occurring, excitation ratiometric, and turn-on fluorescent protein sensor for chloride. Our results show that chloride binding tunes the pK(a) of the chromophore Y66 and shifts the equilibrium from the fluorescent phenolate form to the weakly fluorescent phenol form. The latter likely undergoes excited-state proton transfer to generate a turn-on fluorescence response that is pH-dependent. Moreover, anion selectivity and mutagenesis in the chloride binding pocket provide additional evidence for the proposed chloride sensing mechanism. Given these properties, we anticipate that phiYFP, with further engineering, could be a new tool for imaging cellular chloride dynamics.
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