Detergents modify proteinase K resistance of PrP Sc in different transmissible spongiform encephalopathies (TSEs).
Prion diseases are diagnosed by the detection of their proteinase K-resistant prion protein fragment (PrP(Sc)). Various biochemical protocols use different detergents for the tissue preparation. We found that the resistance of PrP(Sc) against proteinase K may vary strongly with the detergent used. In our study, we investigated the influence of the most commonly used detergents on eight different TSE agents derived from different species and distinct prion disease forms. For a high throughput we used a membrane adsorption assay to detect small amounts of prion aggregates, as well as Western blotting. Tissue lysates were prepared using DOC, SLS, SDS or Triton X-100 in different concentrations and these were digested with various amounts of proteinase K. Detergents are able to enhance or diminish the detectability of PrP(Sc) after proteinase K digestion. Depending on the kind of detergent, its concentration - but also on the host species that developed the TSE and the disease form or prion type - the detectability of PrP(Sc) can be very different. The results obtained here may be helpful during the development or improvement of a PrP(Sc) detection method and they point towards a detergent effect that can be additionally used for decontamination purposes. A plausible explanation for the detergent effects described in this article could be an interaction with the lipids associated with PrP(Sc) that may stabilize the aggregates.
Prion and Dementia Research Unit, Department of Neuropathology, University Medical Center Göttingen, Robert Koch Str. 40, 37075 Göttingen, Germany.
Johanna
J
Breyer
Wiebke M
WM
Wemheuer
Arne
A
Wrede
Catherine
C
Graham
Sylvie L
SL
Benestad
Bertram
B
Brenig
Jürgen A
JA
Richt
Walter J
WJ
Schulz-Schaeffer
D000818:Animals; D002417:Cattle; D003670:Deer; D003840:Deoxycholic Acid / Q000737:chemistry; D003902:Detergents / Q000737:chemistry*; D019286:Endopeptidase K / Q000737:chemistry*; D006801:Humans; D017830:Octoxynol / Q000737:chemistry; D018620:PrPSc Proteins / Q000737:chemistry*; D017096:Prion Diseases / Q000378:metabolism*; D012756:Sheep; D012967:Sodium Dodecyl Sulfate / Q000031:analogs & derivatives / Q000737:chemistry
D003902:Detergents; D018620:PrPSc Proteins; D003840:Deoxycholic Acid; D012967:Sodium Dodecyl Sulfate; D017830:Octoxynol; C408148:sodium laureth sulfate; D019286:Endopeptidase K