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Proteomic analysis of the small intestine and colon epithelia of adenomatous polyposis coli gene‐mutant mice by two‐dimensional gel electrophoresis
Author(s) -
Minowa Takashi,
Ohtsuka Satoshi,
Sasai Hitoshi,
Kamada Masafumi
Publication year - 2000
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/(sici)1522-2683(20000501)21:9<1782::aid-elps1782>3.0.co;2-3
Subject(s) - mutant , microbiology and biotechnology , adenomatous polyposis coli , biology , familial adenomatous polyposis , gene , exon , carcinogenesis , immobilized ph gradient , mutation , gel electrophoresis , mutant protein , amino acid , biochemistry , colorectal cancer , cancer , genetics , enzyme , isoelectric focusing
Mutations of the adenomatous polyposis coli gene (APC) have been implicated in the occurrence of sporadic colon cancer. Various APC mutant strains of mice have been created to better understand the function of this gene. Previously, we had mice express a mutant form of mRNA of the APC protein that encoded 474 amino acids instead of the 2845 amino acids due to exon duplication. These APC mutant mice (APC Δ 474) developed intestinal and mammary tumors, as have other APC mutant mice previously reported (Sasai, H., et al. Carcinogenesis , in press). To elucidate the mechanism of the tumor development, we prepared protein samples from both normal and tumor tissues from APC Δ 474 mutant mice, as well as tissues from normal mice, and used them for proteomic analysis. After two‐dimensional electrophoresis, the gels were silver stained and the protein spots were analyzed. We analyzed about 1000 protein spots per sample and found several protein spots that are specific for normal or tumor samples from APC Δ 474 mutant mice, as well as proteins with altered expression levels. Among the identified protein spots, truncated β‐tubulins were specific to APC Δ 474 mutant mice polyp samples. The apparent molecular mass of these proteins suggested that these β‐tubulins may be truncated very close to the binding site of the anti‐tumor drug taxol.