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United States Department of Agriculture

Agricultural Research Service

Research Project: DETECTION OF TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHY AGENTS IN LIVESTOCK, WILDLIFE, AGRICULTURAL PRODUCTS, AND THE ENVIRONMENT Title: Using synthetic small molecule reagents and antibodies to distinguish among PrP conformers: new uses for old antibodies

Authors
item Silva, Christopher
item Erickson-Beltran, Melissa
item Dynin, Irina
item Carter, John

Submitted to: Meeting Abstract
Publication Type: Abstract Only
Publication Acceptance Date: May 9, 2012
Publication Date: May 12, 2012
Citation: Silva, C.J., Erickson-Beltran, M.L., Dynin, I.A., Carter, J.M. 2012. Using synthetic small molecule reagents and antibodies to distinguish among PrP conformers: new uses for old antibodies. Meeting Abstract. Prion 6(supplement),90.

Technical Abstract: The chemical environment of the amino acids present in PrP is conformation (PrP(Sc) or PrP(c)) dependent. This means that the same amino acid can react differently with the same chemical reagent, depending upon the conformation of that protein. Furthermore, if the site of chemical reaction is the epitiope of an antibody, then the antibody can be used to monitor the extent of this reaction. In order to test this hypothesis, the activated esters small molecules were synthesized and reacted with detergent-solubilized brain extracts from prion-infected mice, uninfected mice, prion-infected hamsters or uninfected hamsters. These reaction mixtures were analyzed by Western blots probed with seven commercially available antibodies. Four of these antibodies recognize an epitope that is encrypted in the PrP(Sc) isoform, but exposed in the PrP(c) isoform. These reagents permit the detection of prion infected brain extracts without the need for proteinase K digestion. In addition these reagents can be used, with an appropriate antibody, to determine which amino acids of PrP(Sc) are exposed on the surface and which are encrypted, thus providing useful structural information. This approach was used to distinguish among strains of hamster-adapted scrapie without the use of proteinase K.

Last Modified: 8/22/2014