The role of glycophosphatidylinositol anchor in the amplification of the scrapie isoform of prion protein in vitro.

Abstract

Transmissible spongiform encephalopathies are associated with an autocatalytic conversion of normal prion protein, PrP(C), to a protease-resistant form, PrPres. This autocatalytic reaction can be reproduced in vitro using a procedure called protein misfolding cyclic amplification (PMCA). Here we show that, unlike brain-derived PrP(C), bacterially-expressed recombinant prion protein (rPrP) is a poor substrate for PrPres amplification in a standard PMCA reaction. The differences between PrP(C) and rPrP appear to be due to the lack of the glycophosphatidylinositol anchor in the recombinant protein. These findings shed a new light on prion protein conversion process and have important implications for the efforts to generate synthetic prions for structural and biophysical studies.