title: Rapid acquisition of beta-sheet structure in the prion protein prior to multimer formation.
abstract: The N-terminally truncated form of the prion protein, PrP 27-30, and the corresponding recombinant protein, rPrP, were solubilized in 0.2% SDS, and the transitions induced by changing the conditions from 0.2% SDS to physiological conditions, i.e. removing SDS, were characterized with respect to solubility, resistance to proteolysis, secondary structure and multimerization. Circular dichroism, electron microscopy and fluorescence correlation spectroscopy were used to study the structural transitions of PrP. Within one minute the alpha-helical structure of PrP was transformed into one that was enriched in beta-sheets and consisted mainly of dimers. Larger oligomers were found after 20 minutes and larger multimers exhibiting resistance to proteolysis were found after several hours. It was concluded that the monomeric alpha-helical conformation was stable in SDS or when attached to the membrane; however, the state of lowest free energy in aqueous solution at neutral pH seems to be the multimeric, beta-sheet enriched conformation.
authors.affiliation: Institut für Physikalische Biologie, Biologisch-Medizinisches Forschungszentrum, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Post
authors.forename: M
authors.initials: M
authors.lastname: Pitschke
authors.forename: O
authors.initials: O
authors.lastname: Schäfer
authors.forename: H
authors.initials: H
authors.lastname: Wille
authors.forename: T R
authors.initials: TR
authors.lastname: Appel
authors.forename: D
authors.initials: D
authors.lastname: Kirsch
authors.forename: I
authors.initials: I
authors.lastname: Mehlhorn
authors.forename: H
authors.initials: H
authors.lastname: Serban
authors.forename: S B
authors.initials: SB
authors.lastname: Prusiner
authors.forename: D
authors.initials: D
authors.lastname: Riesner
mesh_terms: D000818:Animals; D001704:Biopolymers / Q000737:chemistry; D002942:Circular Dichroism; D006224:Cricetinae; D019286:Endopeptidase K / Q000378:metabolism; D007700:Kinetics; D008647:Mesocricetus; D008854:Microscopy, Electron; D011328:Prions / Q000737:chemistry* / Q000378:metabolism; D017433:Protein Structure, Secondary; D013050:Spectrometry, Fluorescence; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D001704:Biopolymers; D011328:Prions; D019286:Endopeptidase K