Development of a novel method for screening of estrogenic compounds using nano-sized bacterial magnetic particles displaying estrogen receptor

Tomoko Yoshino, Fukuichi Kato, Haruko Takeyama, Makoto Nakai, Yoshikuni Yakabe, Tadashi Matsunaga

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

In this study, nano-sized bacterial magnetic particles (BMPs) displaying human estrogen receptor ligand binding domain (ERLBD) on the surface was successfully produced by the magnetic bacterium, Magnetospirillum magneticum AMB-1. Furthermore, a non-isotopic binding assay for estrogenic compounds using the BMPs displaying ERLBD was developed. A BMP membrane-specific protein, Mms16, was used as an anchor molecule to localize ERLBD on the surface of BMPs. ERLBD-BMP complexes were simply extracted by magnetic separation from ruptured AMB-1 transformants and used for the assay based on the competitive binding of alkaline phosphatase conjugated 17β-estradiol (ALP-E2) as a tracer. Dissociation constant of the receptor was 2.3 nM. Inhibition curves were evaluated by the decrease in luminescence intensity resulting from the enzymatic reaction of alkaline phosphatase. The overall simplicity of this receptor binding assay results in a method that can be easily adapted to a high throughput format. Moreover, this method can be integrated into a fully-automated ligand screening system using magnetic separation.

Original languageEnglish
Pages (from-to)105-111
Number of pages7
JournalAnalytica Chimica Acta
Volume532
Issue number2
DOIs
Publication statusPublished - 2005 Mar 14
Externally publishedYes

Fingerprint

estrogenic compound
Estrogen Receptors
ligand
Screening
Estrogens
Ligands
Assays
Magnetic separation
assay
phosphatase
Alkaline Phosphatase
Magnetospirillum
Competitive Binding
luminescence
Luminescence
Anchors
anchor
Estradiol
Bacteria
Membrane Proteins

Keywords

  • alkaline phosphatase conjugated 17β-estradiol (ALP-E2)
  • Bacterial magnetic particles (BMPs)
  • Estrogen receptor ligand binding domain (ERLBD)
  • Estrogenic compounds

ASJC Scopus subject areas

  • Biochemistry
  • Analytical Chemistry
  • Spectroscopy
  • Environmental Chemistry

Cite this

Development of a novel method for screening of estrogenic compounds using nano-sized bacterial magnetic particles displaying estrogen receptor. / Yoshino, Tomoko; Kato, Fukuichi; Takeyama, Haruko; Nakai, Makoto; Yakabe, Yoshikuni; Matsunaga, Tadashi.

In: Analytica Chimica Acta, Vol. 532, No. 2, 14.03.2005, p. 105-111.

Research output: Contribution to journalArticle

Yoshino, Tomoko ; Kato, Fukuichi ; Takeyama, Haruko ; Nakai, Makoto ; Yakabe, Yoshikuni ; Matsunaga, Tadashi. / Development of a novel method for screening of estrogenic compounds using nano-sized bacterial magnetic particles displaying estrogen receptor. In: Analytica Chimica Acta. 2005 ; Vol. 532, No. 2. pp. 105-111.
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