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ARS Home » Pacific West Area » Albany, California » Western Regional Research Center » Produce Safety and Microbiology Research » Research » Publications at this Location » Publication #358151

Research Project: Immunodiagnostics to Detect Prions and Other Important Animal Pathogens

Location: Produce Safety and Microbiology Research

Title: Exploring the conformational differences between PrPSc from classical and atypical scrapie using mass spectrometry

Author
item Silva, Christopher - Chris
item Erickson-Beltran, Melissa
item MARTIN-BURRIEL, INMACULADA - University Of Zaragoza
item BADIOLA, JUAN JOSE - University Of Zaragoza
item REQUENA, JESUS - University Of Santiago De Compostela
item BOLEA, ROSA - University Of Zaragoza

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: 10/11/2018
Publication Date: 10/25/2018
Citation: Silva, C.J., Erickson-Beltran, M.L., Martin-Burriel, I., Badiola, J., Requena, J.R., Bolea, R. 2018. Exploring the conformational differences between PrPSc from classical and atypical scrapie using mass spectrometry. Meeting Abstract. 38(1):65-80. https://doi.org/10.1051/vetres:2006046.
DOI: https://doi.org/10.1051/vetres:2006046

Interpretive Summary:

Technical Abstract: Scrapie propagation is dependent upon compatibility of the PrPSc template with the PrPC substrate and is the molecular basis of the observed “species barrier”. Although atypical scrapie is experimentally transmissible, its epidemiology is consistent with a spontaneous origin. Atypical scrapie is also found in genotypes resistant to classical scrapie (A136R154R171) and can be experimentally transmitted to those resistant genotypes. A multiple reaction monitoring (MRM) mass spectrometry-based method was used to identify and quantitate the peptides derived from the chymotryptic digest of PrPSc isolated from heterozygous sheep naturally infected with classical or atypical scrapie. The intrinsic susceptibility of the specific methionines within these peptides to air oxidation was determined. The MRM approach permitted the determination of the relative amount of each PrP polymorphism present in PrPSc from PrP-heterozygous sheep infected with either classical or atypical scrapie. Furthermore, the total and relative amounts of the PrPC polymorphisms in brain tissue from sheep natural infected with atypical scrapie were determined. The extent of the oxidation of the methionines present in the chymotryptic peptides, M132LGSZMSRPL141 (Z136=A or V) and Y153ZENMY158 (Z154=H or R), was also measured. These approaches provide information on the relative propagation of atypical scrapie PrPSc polymorphisms in the same animals. It also provides information on the relative surface exposure of methionines, based on the extent of their oxidation.