New Structural Approaches to Understanding the Disease Related Forms of the Prion Protein
Abstract
The mouse pron protein peptide (residues 89-143 with the substitution of Leu for Pro atresidue 101) induces prion disease in sensitized mice. Samp%es of this peptide isotope%abe%ed with I 5N were prepared by expression of a fusion in E.co%i c%eaved to yie%d anunmodified peptide and then fibrillized. Hydrogen exchange was allowed to occur in thefibrils for periods from I hour to 6 weeks repeating initial experiments to verify theexchange behavior. The extent of exchange was monitored using peak intensities in I 5N-1 H HSQCnuclear magnetic resonance spectra in DMSO/D2O/TFA solutions. The results are consistent withprevious measurements that we did. Analogous experiments were also done with unlabeledpeptide in fibrils with the analysis done by mass spectroscopy. Fragmentation was improved relative to previous experiments by using a combination of two proteases. With the improvedcoverage it was possible to show that the same protected regions as determined by NMR couldbe identified.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 2007
- Accession Number
- ADA468023
Entities
People
- David E Wemmer
Organizations
- University of California Regents