A High-Throughput Screening Assay of Endocrine-Disrupting Chemicals Using a Receptor-Modified Au-Electrode

Masaharu Murata, Kentaro Yano, Kaori Fukuma, Mizuo Maeda, Yoshiki Katayama

Research output: Contribution to journalArticle

4 Citations (Scopus)

Abstract

There is a great deal of concern that wildlife and human health have been adversely affected by anthropogenic chemicals that can disrupt normal endocrine homeostasis. In order to identify the binding ability of chemicals to human estrogen receptors (hER), we have constructed a biosensor which carries the hER ligand-binding domain on the surface. The receptor was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system, and then the recombinant protein was immobilized on an Au-electrode via Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. This receptor-modified electrode was used to define the binding capacity of several xenoestrogens and anti-estrogen to the hER. The biosensor showed a linear response to the chemicals in a concentration-dependent manner. The order of the detection limits was: 17β-estradiol (10-10 M) > diethylstilbestrol (DES; 10 -8 M), ICI 182780 (10-8 M), dibutyl phthalate (10-8 M), bisphenol A (10-8 M) > p-nonylphenol (10-6 M), testosterone (10-6 M). Compared to the traditional binding assay, our method has the advantage of being more feasible, due to the high-throughput screening assay for evaluating the binding ability of chemicals to the receptors.

Original languageEnglish
Pages (from-to)525-529
Number of pages5
JournalBulletin of the Chemical Society of Japan
Volume77
Issue number3
DOIs
Publication statusPublished - Mar 1 2004

Fingerprint

Endocrine Disruptors
Assays
Screening
Throughput
Estrogen Receptors
Electrodes
Histidine
Biosensors
Nitrilotriacetic Acid
Dibutyl Phthalate
Diethylstilbestrol
Chemisorption
Recombinant Proteins
Sulfhydryl Compounds
Testosterone
Estradiol
Estrogens
Fusion reactions
Health
Ligands

All Science Journal Classification (ASJC) codes

  • Chemistry(all)

Cite this

A High-Throughput Screening Assay of Endocrine-Disrupting Chemicals Using a Receptor-Modified Au-Electrode. / Murata, Masaharu; Yano, Kentaro; Fukuma, Kaori; Maeda, Mizuo; Katayama, Yoshiki.

In: Bulletin of the Chemical Society of Japan, Vol. 77, No. 3, 01.03.2004, p. 525-529.

Research output: Contribution to journalArticle

@article{8f1d37a4285d402ea188be60bd63fae8,
title = "A High-Throughput Screening Assay of Endocrine-Disrupting Chemicals Using a Receptor-Modified Au-Electrode",
abstract = "There is a great deal of concern that wildlife and human health have been adversely affected by anthropogenic chemicals that can disrupt normal endocrine homeostasis. In order to identify the binding ability of chemicals to human estrogen receptors (hER), we have constructed a biosensor which carries the hER ligand-binding domain on the surface. The receptor was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system, and then the recombinant protein was immobilized on an Au-electrode via Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. This receptor-modified electrode was used to define the binding capacity of several xenoestrogens and anti-estrogen to the hER. The biosensor showed a linear response to the chemicals in a concentration-dependent manner. The order of the detection limits was: 17β-estradiol (10-10 M) > diethylstilbestrol (DES; 10 -8 M), ICI 182780 (10-8 M), dibutyl phthalate (10-8 M), bisphenol A (10-8 M) > p-nonylphenol (10-6 M), testosterone (10-6 M). Compared to the traditional binding assay, our method has the advantage of being more feasible, due to the high-throughput screening assay for evaluating the binding ability of chemicals to the receptors.",
author = "Masaharu Murata and Kentaro Yano and Kaori Fukuma and Mizuo Maeda and Yoshiki Katayama",
year = "2004",
month = "3",
day = "1",
doi = "10.1246/bcsj.77.525",
language = "English",
volume = "77",
pages = "525--529",
journal = "Bulletin of the Chemical Society of Japan",
issn = "0009-2673",
publisher = "The Chemical Society of Japan",
number = "3",

}

TY - JOUR

T1 - A High-Throughput Screening Assay of Endocrine-Disrupting Chemicals Using a Receptor-Modified Au-Electrode

AU - Murata, Masaharu

AU - Yano, Kentaro

AU - Fukuma, Kaori

AU - Maeda, Mizuo

AU - Katayama, Yoshiki

PY - 2004/3/1

Y1 - 2004/3/1

N2 - There is a great deal of concern that wildlife and human health have been adversely affected by anthropogenic chemicals that can disrupt normal endocrine homeostasis. In order to identify the binding ability of chemicals to human estrogen receptors (hER), we have constructed a biosensor which carries the hER ligand-binding domain on the surface. The receptor was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system, and then the recombinant protein was immobilized on an Au-electrode via Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. This receptor-modified electrode was used to define the binding capacity of several xenoestrogens and anti-estrogen to the hER. The biosensor showed a linear response to the chemicals in a concentration-dependent manner. The order of the detection limits was: 17β-estradiol (10-10 M) > diethylstilbestrol (DES; 10 -8 M), ICI 182780 (10-8 M), dibutyl phthalate (10-8 M), bisphenol A (10-8 M) > p-nonylphenol (10-6 M), testosterone (10-6 M). Compared to the traditional binding assay, our method has the advantage of being more feasible, due to the high-throughput screening assay for evaluating the binding ability of chemicals to the receptors.

AB - There is a great deal of concern that wildlife and human health have been adversely affected by anthropogenic chemicals that can disrupt normal endocrine homeostasis. In order to identify the binding ability of chemicals to human estrogen receptors (hER), we have constructed a biosensor which carries the hER ligand-binding domain on the surface. The receptor was expressed as an in-frame fusion with ten consecutive histidine residues using a bacterial system, and then the recombinant protein was immobilized on an Au-electrode via Ni(II)-mediated chemisorption using the histidine tag and thiol-modified nitrilotriacetic acid. This receptor-modified electrode was used to define the binding capacity of several xenoestrogens and anti-estrogen to the hER. The biosensor showed a linear response to the chemicals in a concentration-dependent manner. The order of the detection limits was: 17β-estradiol (10-10 M) > diethylstilbestrol (DES; 10 -8 M), ICI 182780 (10-8 M), dibutyl phthalate (10-8 M), bisphenol A (10-8 M) > p-nonylphenol (10-6 M), testosterone (10-6 M). Compared to the traditional binding assay, our method has the advantage of being more feasible, due to the high-throughput screening assay for evaluating the binding ability of chemicals to the receptors.

UR - http://www.scopus.com/inward/record.url?scp=1842531078&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1842531078&partnerID=8YFLogxK

U2 - 10.1246/bcsj.77.525

DO - 10.1246/bcsj.77.525

M3 - Article

VL - 77

SP - 525

EP - 529

JO - Bulletin of the Chemical Society of Japan

JF - Bulletin of the Chemical Society of Japan

SN - 0009-2673

IS - 3

ER -