NR AQQO
AU Yin,S.M.; Sy,M.S.; Po,T.
TI An engineered PrPsc-like molecule from the chimera of mammalian prion protein and yeast Ure2p prion-inducing domain
QU Acta Biochimica et Biophysica Sinica 2004 Feb; 36(2): 128-32
ER Acta Biochim Biophys Sin (Shanghai). 2004 Mar;36(3):176
PT journal article
AB Production of the pathogenic prion isoform PrPsc-like molecules is thought to be useful for understanding the mysterious mechanism of conformational conversion process of prion diseases and proving the "protein-only" hypothesis. In this report, an engineered PrPsc-like conformation was produced from a chimera of mammalian bovine prion protein (bPrP) and yeast Ure2p prion-inducing domain (UPrD). Compared with the normal form of bPrP, the engineered recombinant protein, termed bPrP-UPrD, spontaneously aggregated into ordered fibrils under physiological condition, displaying amyloid-like characteristics, such as fibrillar morphology, birefringence upon binding to Congo red and increased fluorescence intensity with Thioflavine T. Limited resistance to protease K digestion and CD spectroscopy experiments suggested that the structure of bPrP-UPrD had been changed, and adopted a new, high content beta-sheet conformation during the fibrils formation. Moreover, bPrP-UPrD amyloid fibrils could recruit more soluble forms into the aggregates. Therefore, the engineered molecules could mimic significant behaviors of PrPsc and will be helpful for further understanding the mechanism of conformational conversion process.
MH Amyloid/chemistry; Animals; Cattle; Circular Dichroism; Congo Red/pharmacology; Dyes/pharmacology; Electrophoresis, Polyacrylamide Gel; Endopeptidase K/chemistry; Microscopy; Microscopy, Electron; Plasmids/metabolism; PrPsc Proteins/*chemistry/metabolism; Prions/*chemistry; Protein Conformation; Protein Engineering/*methods; Protein Isoforms; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Fusion Proteins/chemistry; Recombinant Proteins/chemistry; Research Support, Non-U.S. Gov't; Saccharomyces cerevisiae/metabolism; Spectrometry, Fluorescence; Thiazoles/chemistry
AD Institute of Microbiology, the Chinese Academy of Sciences, Beijing 100080, China.
SP englisch
PO China