title: Dual conformation of H2H3 domain of prion protein in mammalian cells.
abstract: The concept of prion is applied to protein modules that share the ability to switch between at least two conformational states and transmit one of these through intermolecular interaction and change of conformation. Although much progress has been achieved through the understanding of prions from organisms such as Saccharomyces cerevisiae, Podospora anserina, or Aplysia californica, the criteria that qualify a protein module as a prion are still unclear. In addition, the functionality of known prion domains fails to provide clues to understand the first identified prion, the mammalian infectious prion protein, PrP. To address these issues, we generated mammalian cellular models of expression of the C-terminal two helices of PrP, H2 and H3, which have been hypothesized, among other models, to hold the replication and conversion properties of the infectious PrP. We found that the H2H3 domain is an independent folding unit that undergoes glycosylations and glycosylphosphatidylinositol anchoring similar to full-length PrP. Surprisingly, in some conditions the normally folded H2H3 was able to systematically go through a conversion process and generate insoluble proteinase K-resistant aggregates. This structural switch involves the assembly of amyloid structures that bind thioflavin S and oligomers that are reactive to A11 antibody, which specifically detects protein oligomers from neurological disorders. Overall, we show that H2H3 is a conformational switch in a cellular context and is thus suggested to be a candidate for the conversion domain of PrP.
authors.affiliation: CEA, Institute of Emerging Diseases and Innovative Therapies, SEPIA, 92260 Fontenay-aux-Roses, France.
authors.forename: Zhou
authors.initials: Z
authors.lastname: Xu
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Prigent
authors.forename: Jean-Philippe
authors.initials: JP
authors.lastname: Deslys
authors.forename: Human
authors.initials: H
authors.lastname: Rezaei
mesh_terms: D000682:Amyloid / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D052160:Benzothiazoles; D002460:Cell Line; D006031:Glycosylation; D017261:Glycosylphosphatidylinositols / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D011328:Prions / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D017510:Protein Folding; D055503:Protein Multimerization; D017434:Protein Structure, Tertiary; D011817:Rabbits; D013844:Thiazoles / Q000737:chemistry
chemical_list: D000682:Amyloid; D052160:Benzothiazoles; D017261:Glycosylphosphatidylinositols; D011328:Prions; D013844:Thiazoles; C009462:thioflavin T