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 journalArticlepeer-review

    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

    All Science Journal Classification (ASJC) codes

    • Chemistry(all)

    Fingerprint

    Dive into the research topics of 'A High-Throughput Screening Assay of Endocrine-Disrupting Chemicals Using a Receptor-Modified Au-Electrode'. Together they form a unique fingerprint.

    Cite this