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

10 Citations (Scopus)

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

Fingerprint

proteins
Proteins
probes
sensors
Sensors
bioinstrumentation
Biosensors
mutagenesis
Mutagenesis
cysteine
format
Metal ions
Cysteine
amino acids
Amino acids
Chemical reactions
metal ions
chemical reactions
selectivity
Fluorescence

Keywords

  • Fluorescent probe
  • Fluorescent protein biosensor
  • Mercury sensor
  • Turn-on probe
  • Unnatural amino acid

ASJC Scopus subject areas

  • Bioengineering
  • Fluid Flow and Transfer Processes
  • Process Chemistry and Technology
  • Instrumentation

Cite this

Fluorescent protein-based turn-on probe through a general protection-deprotection design strategy. / Shang, Xin; Wang, Nanxi; Cerny, Ronald; Niu, Wei; Guo, Jiantao.

In: ACS Sensors, Vol. 2, No. 7, 28.07.2017, p. 961-966.

Research output: Contribution to journalArticle

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