title: Prion fibrils of Ure2p assembled under physiological conditions contain highly ordered, natively folded modules.
abstract: The difference between the prion and the non-prion form of a protein is given solely by its three-dimensional structure, according to the prion hypothesis. It has been shown that solid-state NMR can unravel the atomic-resolution three-dimensional structure of prion fragments but, in the case of Ure2p, no highly resolved spectra are obtained from the isolated prion domain. Here, we demonstrate that the spectra of full-length fibrils of Ure2p interestingly lead to highly resolved solid-state NMR spectra. Prion fibrils formed under physiological conditions are therefore well-ordered objects on the molecular level. Comparing the full-length NMR spectra with the corresponding spectra of the prion and globular domains in isolation reveals that the globular part in particular shows almost perfect structural order. The NMR linewidths in these spectra are as narrow as the ones observed in crystals of the isolated globular domain. For the prion domain, the spectra reflect partial disorder, suggesting structural heterogeneity, both in isolation and in full-length Ure2p fibrils, although to different extents. The spectral quality is surprising in the light of existing structural models for Ure2p and in comparison to the corresponding spectra of the only other full-length prion fibrils (HET-s) investigated so far. This opens the exciting perspective of an atomic-resolution structure determination of the fibrillar form of a prion whose assembly is not accompanied by significant conformational changes and documents the structural diversity underlying prion propagation.
authors.affiliation: Institut de Biologie et Chimie des Protéines, UMR 5086 CNRS/Université de Lyon 1, 7 passage du Vercors, 69367 Lyon, France.
authors.forename: Antoine
authors.initials: A
authors.lastname: Loquet
authors.forename: Luc
authors.initials: L
authors.lastname: Bousset
authors.forename: Carole
authors.initials: C
authors.lastname: Gardiennet
authors.forename: Yannick
authors.initials: Y
authors.lastname: Sourigues
authors.forename: Christian
authors.initials: C
authors.lastname: Wasmer
authors.forename: Birgit
authors.initials: B
authors.lastname: Habenstein
authors.forename: Anne
authors.initials: A
authors.lastname: Schütz
authors.forename: Beat H
authors.initials: BH
authors.lastname: Meier
authors.forename: Ronald
authors.initials: R
authors.lastname: Melki
authors.forename: Anja
authors.initials: A
authors.lastname: Böckmann
mesh_terms: D000596:Amino Acids; D003460:Crystallization; D005979:Glutathione Peroxidase; D006358:Hot Temperature; D009682:Magnetic Resonance Spectroscopy; D018583:Pliability; D011328:Prions / Q000737:chemistry*; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D012441:Saccharomyces cerevisiae / Q000737:chemistry*; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry*
chemical_list: D000596:Amino Acids; D011328:Prions; D029701:Saccharomyces cerevisiae Proteins; D005979:Glutathione Peroxidase; C067020:URE2 protein, S cerevisiae