Abstract
We demonstrated a general protection-deprotection strategy for the design of fluorescent protein biosensors through the construction of a turn-on Hg2+ sensor. A combination of fluorescent protein engineering and unnatural amino acid mutagenesis was used. Unlike previously reported fluorescent protein-based Hg2+ sensors that relied on the binding of Hg2+ to the sulfhydryl group of cysteine residues, a well-established chemical reaction, oxymercuration, was transformed into biological format and incorporated into our sensor design. This novel Hg2+ sensor displayed good sensitivity and selectivity both in vitro and in live bacterial cells. Over 60-fold change in fluorescence signal output was observed in the presence of 10 μM Hg2+, while such a change was undetectable when nine other metal ions were tested. This new design strategy could expand the repertoire of fluorescent protein-based biosensors for the detection of small-molecule analytes.
Original language | English (US) |
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Pages (from-to) | 961-966 |
Number of pages | 6 |
Journal | ACS Sensors |
Volume | 2 |
Issue number | 7 |
DOIs | |
State | Published - Jul 28 2017 |
Keywords
- Fluorescent probe
- Fluorescent protein biosensor
- Mercury sensor
- Turn-on probe
- Unnatural amino acid
ASJC Scopus subject areas
- Bioengineering
- Instrumentation
- Process Chemistry and Technology
- Fluid Flow and Transfer Processes