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Volume 24, Number 8—August 2018
Research

Susceptibility of Human Prion Protein to Conversion by Chronic Wasting Disease Prions

Marcelo A. BarriaComments to Author , Adriana Libori, Gordon Mitchell, and Mark W. Head
Author affiliations: National CJD Research and Surveillance Unit, University of Edinburgh, Edinburgh, Scotland, UK (M.A. Barria, A. Libori, M.W. Head); National and OIE Reference Laboratory for Scrapie and CWD, Canadian Food Inspection Agency, Ottawa, Ontario, Canada (G. Mitchell)

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Figure 2

Evaluation of the in vitro conversion of human prion protein (PrP) seeded with the misfolded, disease-associated prion protein form present in chronic wasting disease (CWD)–affected elk brain samples. Western blot analysis for PrP with odd and even number lanes showing reaction mixtures before and after protein misfolding cyclic amplification. A) We incubated 5 elk CWD specimens (elk 0–4) homozygous for Prnp codon 132 methionine (MM) in Tg-HuMM brain substrate (diluted 1:3) and subjected them to

Figure 2. Evaluation of the in vitro conversion of human prion protein (PrP) seeded with the misfolded, disease-associated prion protein form present in chronic wasting disease (CWD)–affected elk brain samples. Western blot analysis for PrP with odd and even number lanes showing reaction mixtures before and after protein misfolding cyclic amplification. A) We incubated 5 elk CWD specimens (elk 0–4) homozygous for Prnp codon 132 methionine (MM) in Tg-HuMM brain substrate (diluted 1:3) and subjected them to a single round of protein misfolding cyclic amplification followed by proteinase K digestion. We performed Western blot analysis by using the mAb 3F4 (for the detection of human protease-resistant prion protein [PrPres]) and 6H4 (detection of CWD PrPres and human PrPres). B) We used a panel of 3 humanized transgenic substrates (Tg-HuMM, Tg-Hu-MV, and Tg-HuVV) to evaluate the susceptibility of the human PrP to conversion. We assessed 3 CWD elk seeds of the132 MM genotype and 2 of the 132 methionine–leucine (ML) genotype. We detected conversion of the human PrP by CWD prions by using the mAb 3F4 after proteinase K digestion. C) We detected total PrPres by using Western blot with mAb 6H4. The elk specimen previously reported (15) is designated elk 0. We performed >5 repeats for the amplification of elk CWD 132 MM seeds and >3 for the 132 ML specimens with similar results. Reference molecular markers have been included. Molecular mass of electrophoretic markers is given. mAb, monoclonal antibody; Tg-HuMM, humanized transgenic PRNP codon 129 homozygous methionine; Tg-HuMV, humanized transgenic methionine/valine; Tg-HuVV, humanized transgenic valine/valine.

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Page created: July 17, 2018
Page updated: July 17, 2018
Page reviewed: July 17, 2018
The conclusions, findings, and opinions expressed by authors contributing to this journal do not necessarily reflect the official position of the U.S. Department of Health and Human Services, the Public Health Service, the Centers for Disease Control and Prevention, or the authors' affiliated institutions. Use of trade names is for identification only and does not imply endorsement by any of the groups named above.
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