Modifiers of prion protein biogenesis and recycling identified by a highly parallel endocytosis kinetics assay.

ABSTRACT: The cellular prion protein, PrP^C, is attached by a glycosylphosphatidylinositol anchor to the outer leaflet of the plasma membrane. Its misfolded isoform PrP^Sc is the causative agent of prion diseases. Conversion of PrP^C into PrP^Sc is thought to take place at the cell surface or in endolysosomal organelles. Understanding the intracellular trafficking of PrP^C may, therefore, help elucidate the conversion process. Here we describe a time-resolved fluorescence energy transfer (FRET) assay reporting membrane expression and real-time internalization rates of PrP^C The assay is suitable for high-throughput genetic and pharmaceutical screens for modulators of PrP^C trafficking. Simultaneous administration of FRET donor and

SUBMITTER: Ballmer BA 

PROVIDER: S-EPMC5437241 | biostudies-literature | 2017 May

REPOSITORIES: biostudies-literature