U.S. flagAn official website of the United States government
                    Here's how you know **Official websites use .gov**
                            
                            A **.gov** website belongs to an official government organization in the United States.
 **Secure .gov websites use HTTPS**
                            
                            A **lock** (                            
                              A locked padlock                                                      
                            
                            ) or **https://** means you’ve safely connected to the .gov website. Share sensitive information only on official, secure websites.


Menu Agricultural Research Service
						
                    U.S. DEPARTMENT OF AGRICULTURE
 Virus and Prion Research: Ames, IA
 close



    Research

                    Research Projects
                                    Publications
                                    Collaborations
                                    News
                People

                    Staff Listing
                                    CareersSearch small
                    Search
            
            ARS Home
                                About ARS
                                Contact Us




ARS Home Midwest Area Ames, Iowa National Animal Disease Center Virus and Prion Research Research Publications at this Location





- ARS National Programs
- Search for a scientific manuscript
- Search for a research project
- ARS Software
- Programs and Projects
- Collaborations
- Subjects of Investigation
- Publications

Title:   Author  item
                                            Vrentas, Catherineitem
                                            Greenlee, Justinitem
                                            BARON, THIERRY - French Agency Of Food Security And Sanitationitem
                                            CARAMELLI, MARIA - Institute Of Experimental Zooprofilatticitem
                                            CZUB, STEFANIE - Aafc Lethrdge Research Centeritem
                                            Nicholson, Eric
  Submitted to: BioMed Central (BMC) Veterinary Research
                                    Publication Type: Peer Reviewed Journal
                                    Publication Acceptance Date: 8/12/2013
                                    Publication  Date: 8/15/2013
                                    Citation: Vrentas, C.E., Greenlee, J.J., Baron, T., Caramelli, M., Czub, S., Nicholson, E.M. 2013. Stability properties of PrPSc from cattle with experimental transmissible spongiform encephalopathies: use of a rapid whole homogenate, protease-free assay. BMC Veterinary Research. 9:167.Interpretive Summary: Transmissible spongiform encephalopathies (TSEs), including scrapie in sheep, chronic wasting disease in cervids, and bovine spongiform encephalopathy (BSE) in cattle, are fatal diseases of the nervous system associated with the accumulation of a misfolded form of the prion protein (PrP**Sc). Different strains of BSE exist, including classical (infectious) BSE and atypical BSE (the H-type and L-type strains, proposed to arise spontaneously; and E211K BSE, an inherited form). TSE strains can be differentiated by a combination of features including a stability assay that measures the unfolding of PrP**Sc. In this work, we provide the first direct application of the stability assay to strains of BSE (as well as other cattle-passaged TSEs) in order to assess potential uses of this assay in strain discrimination as well as to improve our understanding of the properties and relationships of these disease strains. Overall, we demonstrate that the stability of cattle PrP**Sc is largely invariant with the exception of H-type BSE, which exhibits a higher stability. The stability of E211K BSE is consistent with its previous assignment as an H-type strain. While the difference in stability under these experimental conditions is not large enough for rapid use in the diagnostic laboratory, future research on the role of sample conditions in this assay may be able to utilize these differences for assay development. In addition, our results highlight the need for continuing research on the biochemical nature of BSE strains, in order to the understand the relationships between stability properties and other assays used to discriminate between strains, as well as to understand the origin of tissue-specific differences in PrP**Sc.
                                Technical Abstract: Transmissible Spongiform Encephalopathies (TSEs), including scrapie in sheep, chronic wasting disease (CWD) in cervids, and bovine spongiform encephalopathy (BSE), are fatal diseases of the nervous system associated with accumulation of misfolded prion protein (PrPSc). Different strains of BSE exist, including classical BSE, H- and L-type atypical BSE, and H-type BSE associated with the inherited E211K prion protein polymorphism. Strains are associated with different PrPSc conformations that can be probed by the stability assay, in which PrPSc is treated with increasing concentrations of the denaturant guanidine hydrochloride (GdnHCl). Here, we provide the first comprehensive application of the stability assay to brain tissue from cattle experimentally infected with various TSE isolates. Consistent with previous findings from a single Japanese isolate, the L-type form of BSE is not distinguishable from classical BSE in this assay. In contrast, H-type BSE, as well as E211K BSE, exhibits higher stability than classical BSE, suggesting that increased protection against protease digestion at the BSE N-terminus is associated with a higher stability in GdnHCl. While the difference in stability is not large enough in our version of the assay for effective use in a diagnostic laboratory setting, the use of alternative experimental conditions may enhance this effect. TSEs from other natural host species that have been passaged in cattle, including CWD and transmissible mink encephalopathy, were not distinguishable from cattle classical BSE. These results suggest that the core of PrPSc, as probed in this assay, has similar stability properties among cattle-passaged TSE strains and that the conformational differences that lead to changes in the proteinase K cleavage site do not cause large changes in the stability of PrPSc from BSE-affected cattle.


Return to top


Connect with ARS Sign up


ARS Home
                            USDA.gov
                            Plain Writing
                            Policies & Links
                            Civil Rights Statements
                            FOIA
                            Accessibility Statement
                            Privacy Policy
                            Non-Discrimination Statement
                            Quality of Information
                            USA.gov
                            WhiteHouse.gov