title: Purification and fibrillation of full-length recombinant PrP.
abstract: Misfolding and aggregation of prion protein (PrP) is related to several neurodegenerative diseases in humans such as Creutzfeldt-Jacob disease, fatal familial insomnia, and Gerstmann-Straussler-Sheinker disease. Certain applications in prion area require recombinant PrP of high purity and quality. Here, we report an experimental procedure for expression and purification of full-length mammalian PrP. This protocol has been proved to yield PrP of extremely high purity that lacks PrP adducts, which are normally generated as a result of spontaneous oxidation or degradation. We also describe methods for the preparation of amyloid fibrils from recombinant PrP in vitro. Recombinant PrP fibrils can be used as a noninfectious synthetic surrogate of PrP(Sc) for development of prion diagnostics including the generation of PrP(Sc)-specific antibody.
authors.affiliation: Center for Biomedical Engineering and Technology, Department of Anatomy and Neurobiology, University of Maryland, Baltimore, MD, USA.
authors.forename: Natallia
authors.initials: N
authors.lastname: Makarava
authors.forename: Ilia V
authors.initials: IV
authors.lastname: Baskakov
mesh_terms: D000682:Amyloid / Q000737:chemistry; D000818:Animals; D005591:Chemical Fractionation / Q000379:methods*; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004926:Escherichia coli / Q000166:cytology / Q000235:genetics; D002479:Inclusion Bodies / Q000378:metabolism; D051379:Mice; D008856:Microscopy, Fluorescence; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D055503:Protein Multimerization; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification*; D012995:Solubility; D014169:Transformation, Bacterial
chemical_list: D000682:Amyloid; D011328:Prions; D011994:Recombinant Proteins