Novel method for evaluation of chemicals based on ligand-dependent recruitment of GFP labeled coactivator to estrogen receptor displayed on bacterial magnetic particles

Tomoko Yoshino, Chihiro Kaji, Makoto Nakai, Fumiyo Saito, Haruko Takeyama, Tadashi Matsunaga

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

We established a novel method to evaluate endocrine disrupting chemicals (EDCs) by assembling the estrogen receptor-ligand binding domain (ERLBD) and GFP labeled coactivator on magnetic nanoparticles. EDC can promote or inhibit coactivator recruitment to the ligand-ERLBD complex. ERLBD was displayed on the surface of nano-sized bacterial magnetic particles (BacMPs) produced by the magnetic bacterium, Magnetospirillum magneticum AMB-1. Our method resulted in 38 molecules of ERLBD molecules on a BacMPs with diameter of 75 nm. Furthermore, ligand-dependent recruitment assays of GFP labeled coactivator to ERLBD-BacMPs was performed by measuring the fluorescence intensity. 17β-estradiol (E2), estriol, diethylstilbestrol, zeralenone (full agonist), octylphenol (partial agonist) and ICI 182,780 (antagonist) were evaluated by this method. Full agonists tested showed increased fluorescence with increasing agonist concentration. Octylphenol had lower fluorescence intensity than E2. ICI 182,780 did not produce any fluorescence. The method developed in this study can evaluate the estrogenic potential of chemicals by discriminating whether they are an ER full agonist, partial agonist, or antagonist. Finally, this method is amenable adaptation into a high throughput format by using automated magnetic separation.

Original languageEnglish
Pages (from-to)71-77
Number of pages7
JournalAnalytica Chimica Acta
Volume626
Issue number1
DOIs
Publication statusPublished - 2008 Sep 19
Externally publishedYes

Fingerprint

Estrogen Receptors
ligand
Ligands
fluorescence
Fluorescence
Endocrine Disruptors
Magnetospirillum
Magnetic separation
Estriol
Molecules
Diethylstilbestrol
evaluation
method
chemical
particle
Nanoparticles
Estradiol
Assays
Bacteria
Throughput

Keywords

  • Bacterial magnetic particles
  • Coactivator
  • Estrogen receptor
  • Ligand

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

Cite this

Novel method for evaluation of chemicals based on ligand-dependent recruitment of GFP labeled coactivator to estrogen receptor displayed on bacterial magnetic particles. / Yoshino, Tomoko; Kaji, Chihiro; Nakai, Makoto; Saito, Fumiyo; Takeyama, Haruko; Matsunaga, Tadashi.

In: Analytica Chimica Acta, Vol. 626, No. 1, 19.09.2008, p. 71-77.

Research output: Contribution to journalArticle

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