Quantitative Analysis of Prion Protein by Immunoblotting

Kaori Takekida; Yutaka Kikuchi; Takeshi Yamazaki; Motohiro Horiuchi; Tomoshi Kakeya; Morikazu Shinagawa; Kosuke Takatori; Akio Tanimura; Ken-ichi Tanamoto; Jun-ichi Sawada

doi:10.1248/jhs.48.288

RESEARCH LETTERS

Quantitative Analysis of Prion Protein by Immunoblotting

Kaori Takekida, Yutaka Kikuchi, Takeshi Yamazaki, Motohiro Horiuchi, Tomoshi Kakeya, Morikazu Shinagawa, Kosuke Takatori, Akio Tanimura, Ken-ichi Tanamoto, Jun-ichi Sawada

Author information

Kaori Takekida
Showa Woman's University
Yutaka Kikuchi
Division of Microbiology, National Institute of Health Sciences
Takeshi Yamazaki
Division of Food Additives, National Institute of Health Sciences
Motohiro Horiuchi
Research Center for Protozoan, Department of Veterinary Medicine, Obihiro University of Agriculture and Veterinary medicine
Tomoshi Kakeya
Division of Microbiology, National Institute of Health Sciences
Morikazu Shinagawa
Laboratory of Veterinary Public Health, Department of Veterinary Medicine, Obihiro University of Agriculture and Veterinary medicine
Kosuke Takatori
Division of Microbiology, National Institute of Health Sciences
Akio Tanimura
Showa Woman's University
Ken-ichi Tanamoto
Division of Microbiology, National Institute of Health Sciences
Jun-ichi Sawada
Division of Biochemistry and Immunochemistry, National Institute of Health Sciences

Keywords: prion protein, immunoblotting, quantification, bovine, PrP

JOURNAL FREE ACCESS

2002 Volume 48 Issue 3 Pages 288-291

DOI https://doi.org/10.1248/jhs.48.288

Details

Abstract

Transmissible spongiform encephalopathy (TSE) is a neurodegenerative disease characterized by spongiform degeneration and accumulation of an infectious isoform (PrP^Sc) of the prion protein in the central nervous system. PrP^Sc originates from a ubiquitous cellular prion protein (PrP^C). We attempted to develop an easy method of quantitative analysis of PrP by immunoblotting based on densitometry data for PrP bands in immunoblots. Both PrP^C and PrP^Sc yield three bands in immunoblots, and they correspond to PrP molecules carrying two, one, and no Asn-linked sugar chains. We used bovine PrP^C 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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