Quantitative Analysis of Prion Protein by Immunoblotting

抄録

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.