NR ASYD
AU Vendrely,C.; Valadie,H.; Bednarova,L.; Cardin,L.; Pasdeloup,M.; Cappadoro,J.; Bednar,J.; Rinaudo,M.; Jamin,M.
TI Assembly of the full-length recombinant mouse prion protein I. Formation of soluble oligomers.
QU Biochimica et Biophysica Acta 2005 Aug 5; 1724(3): 355-66
PT journal article
AB The conversion of a monomeric alpha-helix-rich isoform to multimeric beta-sheet-rich isoforms is a prominent feature of the conversion between PrPc and PrPsc. We mimicked this process in vitro by exposing an unglycosylated recombinant form of the full-length mouse prion protein ((Mo)PrP(23-231)) to an acidic pH, at 37 degrees C, and we monitored the kinetics of conformational change and assembly. In these conditions, monomeric (Mo)PrP(23-231) converts slowly to two ensembles of soluble oligomers that are separated by size exclusion chromatography. The larger oligomers (I) are unstable, and their formation involves almost no change in secondary structure content. The smaller oligomers (II) form stable spherical or annular particles containing between 8 and 15 monomers as determined by multi-angle laser light scattering (MALLS). Their formation is concomitant with the main, thought limited, change in the secondary structure content (10%) seen by Fourier Transform Infrared (FTIR) spectroscopy. Even if these oligomers conserve a large part of the secondary structure of monomeric PrP, they exhibit amyloid features with the appearance of intermolecular beta-structure as revealed by the appearance of an IR band below 1620 cm(-1).
MH Animals; Chromatography, Gel; Circular Dichroism; Heat; Hydrogen-Ion Concentration; Kinetics; Mice; Peptide Fragments/chemistry/metabolism/ultrastructure; Prions/chemistry/genetics/*metabolism; Protein Denaturation; Protein Isoforms/chemistry/metabolism/ultrastructure; *Protein Processing, Post-Translational; Recombinant Proteins/chemistry/*metabolism/ultrastructure; Research Support, Non-U.S. Gov't; Solubility; Spectrophotometry, Ultraviolet; Spectroscopy, Fourier Transform Infrared
AD Laboratoire de Biophysique Moleculaire et Cellulaire, Universite Joseph Fourier, BMC/DRDC, 17 rue des Martyrs, 38054 Grenoble Cedex 9, France.
SP englisch
PO Niederlande