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J.Health Sci., 50(3), 257-263, 2004
Isolation of Genes Regulated by Peroxisome Proliferator-Activated Receptor
gamma (PPARgamma) by Two-Dimensional Electrophoresis and Mass Spectrometry
Saori Tsuchiya,a Emi
Arimoto,a, b Makoto
Nishizuka,a Ryo
Taguchi,a 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 and
bLaboratory of Environmental Biochemistry, Graduate School of Pharmaceutical Sciences,
Osaka University, 1-6 Yamada-oka, Suita, Osaka 565-0871, Japan
To isolate the genes regulated by peroxisome proliferator-activated receptor
gamma (PPARgamma), we first developed a stable transformant expressing
PPARgamma. Using cytosol and nuclear fractions prepared from this cell line, we
performed two-dimensional electrophoresis. A comparison of the electrophoretic patterns with those from control cells
revealed many spots to be up-regulated or down-regulated proteins. Some spots were subjected to mass
spectrometric analyses, and triosephosphate isomerase and galectin-3 were identified as proteins down-regulated by
PPARgamma. Moreover, Northern blot analyses revealed that the expression level of both genes decreased in
PPARgamma-expressing cells. These results strongly suggest that triosephosphate isomerase and galectin-3 are target genes whose expression
is mediated by PPARgamma.
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