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J.Health Sci., 50(1), 25-32, 2004
Development of a Competitive Enzyme Immunoassay for Detection of Capacity
of Chemicals to Bind Quail Estrogen
Receptor alpha and beta
Shinobu Maekawa,a Makoto
Nishizuka,a Shiro
Heitaku,a Masaaki
Kunimoto,a
Jun-ichi Nishikawa,b Kouhei
Ichikawa,c Kiyoshi
Shimada,c and Masayoshi
Imagawa*, a
aDepartment of Molecular Biology, Graduate School of Pharmaceutical Sciences, Nagoya City University, 3-1 Tanabe-dori,
Mizuho-ku, Nagoya, Aichi 467-8603, Japan,
bLaboratory of Environmental Biochemistry, Graduate School of Pharmaceutical Sciences,
Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan, and
cLaboratory of Animal Physiology, Graduate School of
Bioagricultural Sciences, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, Aichi 464-8601, Japan
In vitro binding assays are useful in the initial screening of endocrine disrupting chemicals. Such assays
should be applied to the estrogen receptors (ER) of not only humans but also wildlife. As a system for birds is yet
to established, we expressed the ligand binding domain (LBD) of quail
ER alpha and ER beta as a fusion protein with
glutathione S-transferase and using these proteins, developed two systems (a competitive enzyme immunoassay
and a fluorescence polarization assay) for assaying the capacity to bind ERs
in vitro. Moreover, 20 test chemicals selected by Ministry of the Environment of Japan were evaluated in terms of binding ability. Both systems
worked well, the competitive enzyme immunoassay proving especially powerful, since it needs no special
equipment. This system is applicable to other species including fish, amphibians and reptiles when information on the
LBD of ER is available.
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