Prion protein NMR structures of cats, dogs, pigs, and sheep

Abstract

The NMR structures of the recombinant cellular form of the prion proteins (PrP C ) of the cat (Felis catus), dog (Canis familiaris), and pig (Sus scrofa), and of two polymorphic forms of the prion protein from sheep (Ovis aries) are presented. In all of these species, PrP C consists of an N-terminal flexibly extended tail with Ϸ100 amino acid residues and a C-terminal globular domain of Ϸ100 residues with three ␣-helices and a short antiparallel ␤-sheet. Although this global architecture coincides with the previously reported murine, Syrian hamster, bovine, and human PrP C structures, there are local differences between the globular domains of the different species. Because the five newly determined PrP C structures originate from species with widely different transmissible spongiform encephalopathy records, the present data indicate previously uncharacterized possible correlations between local features in PrP C threedimensional structures and susceptibility of different mammalian species to transmissible spongiform encephalopathies.mammalian species ͉ feline transmissible spongiform encephalopathy ͉ scrapie T he prion protein (PrP) in mammalian organisms has attracted keen interest because of its relation to a group of invariably fatal neurodegenerative diseases, the transmissible spongiform encephalopathies (TSEs) or ''prion diseases,'' which include bovine spongiform encephalopathy (BSE), CreutzfeldtJakob disease in humans, feline spongiform encephalopathy, and scrapie in sheep. It is well established that expression of the host-encoded PrP is essential for TSE propagation (1, 2). In transgenic mice lacking the gene that encodes PrP, TSEs could not be observed, and the susceptibility toward TSE of these mice could only be restored by reestablishing PrP expression (3). High sequence conservation of PrP in mammalian species (4) indicates that this protein is functionally important in the healthy organism (1, 2), but the search for this unknown function is still ongoing.PrP was identified in the context of TSEs in an aggregated ''scrapie'' isoform of PrP (PrP Sc ) (5), which copurifies with the infective agent (6). This osbservation, the apparent stability of the infectious agent under DNA͞RNA denaturing conditions (7), and the unusual progression of the disease (8) led to the ''protein-only hypothesis.'' This hypothesis proposes that the major component, if not the only one, of the infectious particle causing TSE is a protein, i.e., presumably PrP Sc (1,(7)(8)(9)). An early observation in TSE infections has been the species barrier (10). Compared with transmission with infectious material from the same species, the incubation time for onset of TSEs is prolonged if a given species is challenged with infectious brain homogenate originating from another species. The incubation time may be reduced by consecutive passages within the new host, whereby the adaptation to the new host can take several generations for the disease to show clinical signs (11). In vivo and in vitro experiments indicated that the species ba...

Molecular Dynamics as an Approach to Study Prion Protein Misfolding and the Effect of Pathogenic Mutations

“…For PrP, the first [8] and most abundant structural information has been obtained by nuclear magnetic resonance techniques (NMR), which has resulted in experimentally derived models of PrP of a wide variety of species [64][65][66][67][68][69][70]. For human and sheep PrP, structures determined by X-ray crystallography have also been reported, both with and without antibodies bound [71][72][73][74][75].…”

A prion disease of cervids: Chronic wasting disease

“…The PrP C structure has been solved by nuclear magnetic resonance (NMR) analysis from recombinant prion protein from a library of vertebrate species [13,21,31,[45][46][47]63]. The global architecture of the various mammalian PrP structures are nearly identical.…”

Canine MDCK cell lines are refractory to infection with human and mouse prions

“…After washing, the plate was incubated with an anti-mouse IgG1-HRP secondary antibody (Zymed, 1mg/mL, 1:18000) and the reaction was visualized with a chromogenic substrate as described above. Bacterially were produced and purified as described previously [15].…”