A new fluorescence complementation biosensor for detection of estrogenic compounds

Michael J. Mclachlan, John A. Katzenellenbogen, Huimin Zhao

Research output: Contribution to journalArticlepeer-review

Abstract

Estrogenic compounds are an important class of hormonal substances that can be found as environmental contaminants, with sources including pharmaceuticals, human and animal waste, the chemical industry, and microbial metabolism. Here we report the creation of a biosensor useful for monitoring such compounds, based on complementation of fluorescent protein fragments. A series of sensors were made consisting of fragments of a split mVenus fluorescent protein fused at several different N-terminal and C-terminal positions flanking the ligand binding domain of the estrogen receptor alpha. When expressed in HeLa cells, sensor 6 (ERα 312-595) showed a nine-fold increase in fluorescence in the presence of estrogen receptor agonists or antagonists. Sensor 2 (ERα 281-549) discriminated between agonists and antagonists by showing a decrease in fluorescence in the presence of agonists while being induced by antagonists. The fluorescent signal of sensor 6 increased over a period of 24h, with a two-fold induction visible at 4h and four-fold at 8h of ligand incubation. Ligand titration showed a good correlation with the known relative binding affinities of the compound. The sensor could detect a number of compounds of interest that can act as environmental endocrine disruptors. The lack of a substrate requirement, the speed of signal development, the potential for high throughput assays, and the ability to distinguish agonists from antagonists make this an attractive sensor for widespread use.

Original languageEnglish (US)
Pages (from-to)2794-2803
Number of pages10
JournalBiotechnology and bioengineering
Volume108
Issue number12
DOIs
StatePublished - Dec 2011

Keywords

  • Estrogen receptor
  • Estrogenic compounds
  • Fluorescence complementation
  • Molecular biosensor
  • Split proteins

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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