Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II)

• The GFP chemosensors performed a fluorescence light-up response to Hg(II).
• The underlying mechanism was unveiled as a shift between two ground-state populations of GFP.
• The light-up imaging of Hg(II) in living cells was achieved.

Hg(II) is well-known for quenching fluorescence in a distance dependent manner. Nevertheless, when we exposed the fluorophore of a green fluorescent protein (GFP) toward Hg(II), through H148C mutation, the GFP fluorescence could be “lighted up” by Hg(II) down to sub-nM level. The detection linear range is 0.5–3.0 nM for protein solutions at 8.0 nM. The GFPH148C protein displayed a promising selectivity toward Hg(II) and also the cellular imaging capacity. Spectra measurements suggested that the ground-state redistribution of protein contributed to the fluorescence enhancement, which was found not limited to Hg(II), and thus presented an opening for building a pool of GFP-based chemosensors toward other heavy metal ions.

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