Fluorescent protein-based turn-on probe through a general protection-deprotection design strategy

Xin Shang, Nanxi Wang, Ronald Cerny, Wei Niu, Jiantao Guo

Research output: Contribution to journalArticle

17 Scopus citations

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 languageEnglish (US)
Pages (from-to)961-966
Number of pages6
JournalACS Sensors
Volume2
Issue number7
DOIs
StatePublished - 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

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