NR ALHL
AU Swietnicki,W.; Morillas,M.; Chen,S.G.; Gambetti,P.; Surewicz,W.K.
TI Aggregation and fibrillization of the recombinant human prion protein huPrP90-231
QU Biochemistry 2000 Jan 18; 39(2): 424-31
PT journal article
AB According to the "protein-only" hypothesis, the critical step in the pathogenesis of prion diseases is the conformational transition between the normal (PrPc) and pathological (PrPsc) isoforms of prion protein. To gain insight into the mechanism of this transition, we have characterized the biophysical properties of the recombinant protein corresponding to residues 90-231 of the human prion protein (huPrP90-231). Incubation of the protein under acidic conditions (pH 3.6-5) in the presence of 1 M guanidine-HCl resulted in a time-dependent transition from an alpha-helical conformation to a beta-sheet structure and oligomerization of huPrP90-231 into large molecular weight aggregates. No stable monomeric beta-sheet-rich folding intermediate of the protein could be detected in the present experiments. Kinetic analysis of the data indicates that the formation of beta-sheet structure and protein oligomerization likely occur concomitantly. The beta-sheet-rich oligomers were characterized by a markedly increased resistance to proteinase K digestion and a fibrillar morphology (i.e., they had the essential physicochemical properties of PrPsc). Contrary to previous suggestions, the conversion of the recombinant prion protein into a PrPsc-like form could be accomplished under nonreducing conditions, without the need to disrupt the disulfide bond. Experiments in urea indicate that, in addition to acidic pH, another critical factor controlling the transition of huPrP90-231 to an oligomeric beta-sheet structure is the presence of salt.
MH Chromatography, Gel; Circular Dichroism; Comparative Study; Endopeptidase K; Guanidine; Human; Hydrogen-Ion Concentration; Magnetic Resonance Spectroscopy; Peptide Fragments/*chemistry; Plasmids; PrPc Proteins/chemistry; PrPsc Proteins/chemistry; Prions/*chemistry; Protein Conformation; Recombinant Proteins/chemistry; Sodium Chloride; Support, U.S. Gov't, P.H.S.; Urea
AD Department of Pathology, Case Western Reserve University, Cleveland, Ohio 44106, USA
SP englisch
PO USA