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J.Health Sci., 48(3), 288-291, 2002
Quantitative Analysis of Prion Protein by Immunoblotting
Kaori Takekida,a Yutaka Kikuchi,b Takeshi Yamazaki,*, c Motohiro Horiuchi,d, e Tomoshi Kakeya,b Morikazu Shinagawa,e Kosuke Takatori,b Akio Tanimura,a Ken-ichi Tanamoto,b and Jun-ichi Sawadaf
aShowa Womanﾕs University, 1-7 Taishido, Setagaya, Tokyo 154-8533, Japan, bDivision of Microbiology, cDivision of Food Additives, and fDivision of Biochemistry and Immunochemistry, National Institute of Health Sciences, 1-18-1 Kamiyoga, Setagaya, Tokyo 158-8501, Japan, dResearch Center for Protozoan, and eLaboratory of Veterinary Public Health, Department of Veterinary Medicine, Obihiro University of Agriculture and Veterinary medicine, Inada-cho, Obihiro, Hokkaido 080-8555, Japan
Transmissible spongiform encephalopathy (TSE) is a neurodegenerative disease characterized by spongiform degeneration and accumulation of an infectious isoform (PrPSc) of the prion protein in the central nervous system. PrPSc originates from a ubiquitous cellular prion protein (PrPC). We attempted to develop an easy method of quantitative analysis of PrP by immunoblotting based on densitometry data for PrP bands in immunoblots. Both PrPC and PrPSc yield three bands in immunoblots, and they correspond to PrP molecules carrying two, one, and no Asn-linked sugar chains. We used bovine PrPC as a model protein in the immunoblotting study. We removed the Asn-linked sugar chains from the PrP molecules with N-glycanase to convert all three glycoforms of PrP into a single band of the deglycosylated form and determined the PrP by densitometry calibrated with recombinant bovine PrP.
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