title: Prion rods contain an inert polysaccharide scaffold.
abstract: A polysaccharide consisting of mainly 1,4-linked glucose units was found associated with prion rods, which are composed mainly of insoluble aggregates of the N-terminally truncated prion protein (PrP 27-30) exhibiting the ultrastructural and tinctorial properties of amyloid. The polysaccharide differs in composition from the Asn-linked oligosaccharides and the GPI-anchor of the prion protein. Prion rods were prepared from scrapie-infected hamster brains using two different purification protocols. Prolonged digestion of rods with proteinase K reduced PrP by a factor of at least 500, leaving about 10% (w/w) of the sample as an insoluble remnant. Only glucose was obtained by acid hydrolysis of the remnant and methylation analysis showed 80% 1,4-, 15% 1,6- and 5% 1,4,6-linked glucose units. The physical and chemical properties as well as the absence of terminal glucose units indicate a very high molecular mass of the polysaccharide. No evidence was found for covalent bonds between PrP and the polysaccharide. The polysaccharide certainly contributes to the unusual chemical and physical stability of prion rods, acting like a scaffold. A potential structural and/or functional relevance of the polysaccharide scaffold is discussed.
authors.affiliation: Institut für Physikalische Biologie, Biologisch-Medizinisches Forschungszentrum, Heinrich-Heine-Universität Düsseldorf, Germany.
authors.forename: T R
authors.initials: TR
authors.lastname: Appel
authors.forename: C
authors.initials: C
authors.lastname: Dumpitak
authors.forename: U
authors.initials: U
authors.lastname: Matthiesen
authors.forename: D
authors.initials: D
authors.lastname: Riesner
mesh_terms: D000818:Animals; D001216:Asparagine / Q000737:chemistry; D001671:Biochemistry / Q000379:methods; D002236:Carbohydrate Conformation; D002240:Carbohydrate Sequence; D006224:Cricetinae; D019286:Endopeptidase K / Q000737:chemistry / Q000378:metabolism; D005936:Glucans / Q000032:analysis / Q000737:chemistry; D005947:Glucose / Q000032:analysis; D017261:Glycosylphosphatidylinositols / Q000737:chemistry; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D011328:Prions / Q000737:chemistry* / Q000378:metabolism
chemical_list: D005936:Glucans; D017261:Glycosylphosphatidylinositols; D011134:Polysaccharides; D011328:Prions; D001216:Asparagine; D019286:Endopeptidase K; D005947:Glucose