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. Samples of this peptide, isotope labeled with 15N, have been prepared by expression of a fusion in E.coli, cleaved to yield anunmodified peptide, and then fibrillized. Hydrogen exchange was allowed to occur in the fibrils for periods from 1 hour to 6 weeks. The extent of exchange was monitored using peak intensities in 15N-1H HSQC nuclear magnetic resonance spectra in DMSO/D2O/TFA solutions that had previously been assigned. The exchange data show a high fraction of amides are protected,with a core set (residues 104-109 and 117-135) exchanging very slowly (<10% exchange in 6 weeks). Some biphasic exchange was observed for residues between 110 and 116 suggesting aconformationally heterogeneous region, possibly relating to strain behavior. Samples of unlabeled, uniformly 13C/15N labeled and selectively 13C labeled peptide were prepared for solid state NMR experiments.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jul 01, 2006
- Accession Number
- ADA468058
Entities
People
- David E Wemmer
Organizations
- University of California Regents