title: Expression and characterisation of fully posttranslationally modified cellular prion protein in Pichia pastoris.
abstract: Prion diseases are fatal neurodegenerative diseases which occur as sporadic, genetic, and transmissible disorders. A molecular hallmark of prion diseases is the conformational conversion of the host-encoded cellular form of the prion protein (PrPC) into its misfolded pathogenic isoform (PrPSc). PrPSc is the main component of the pathological and infectious prion agent. The study of the conversion mechanism from PrPC to PrPSc is a major field in prion research. PrPC is glycosylated and attached to the plasma membrane via its glycosyl phosphatidyl inositol (GPI)-anchor. In this study we established and characterised the expression of fully posttranslationally modified mammalian Syrian golden hamster PrPC in the yeast Pichia pastoris using native PrPC-specific N- and C-terminal signal sequences. In vivo as well as in vitro-studies demonstrated that the signal sequences controlled posttranslational processing and trafficking of native PrPC, resulting in PrPC localised in the plasma membrane of P. pastoris. In addition, the glycosylation pattern of native PrPC could be confirmed.
authors.forename: Jendrik
authors.initials: J
authors.lastname: Marbach
authors.forename: Peter
authors.initials: P
authors.lastname: Zentis
authors.forename: Philipp
authors.initials: P
authors.lastname: Ellinger
authors.forename: Henrik
authors.initials: H
authors.lastname: Müller
authors.forename: Eva
authors.initials: E
authors.lastname: Birkmann
mesh_terms: D000818:Animals; D002461:Cell Line, Transformed; D006224:Cricetinae; D005822:Genetic Vectors; D006031:Glycosylation; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D008647:Mesocricetus; D010843:Pichia / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D018620:PrPSc Proteins / Q000737:chemistry* / Q000235:genetics* / Q000378:metabolism; D011499:Protein Processing, Post-Translational / Q000235:genetics; D021382:Protein Sorting Signals / Q000235:genetics; D021381:Protein Transport / Q000235:genetics; D014162:Transfection
chemical_list: D018620:PrPSc Proteins; D021382:Protein Sorting Signals; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins