title: Structural differences between TSEs strains investigated by FT-IR spectroscopy.
abstract: Strain diversity in transmissible spongiform encephalopathies (TSEs) has been suggested to be "enciphered" in the structure of the misfolded prion protein isoform PrP(Sc). We have recently demonstrated the strain typing potential of the FT-IR spectroscopy technique, analyzing four different TSE agents adapted to Syrian hamsters [A. Thomzig, S. Spassov, M. Friedrich, D. Naumann and M. Beekes, Discriminating scrapie and BSE isolates by infrared spectroscopy of pathological prion protein J. Biol. Chem. 279 (2004) 33847-33854.] [1]. In the present paper, we have extended the FT-IR study, exploring the secondary structure, temperature stability, and hydrogen-deuterium exchange characteristics of PrP27-30, from the TSE agents 263K, ME7-H, 22A-H, and BSE-H. The strain differentiation capacity of the FT-IR approach was objectively proven for the first time by multivariate cluster analysis. The second derivative FT-IR spectra obtained from dried protein films or samples hydrated in H(2)O or D(2)O consistently exhibited strain-specific infrared characteristics in the secondary structure sensitive amide I region, complemented by strain dependent spectral traits in the amide II and amide A absorption regions, and the different H/D-exchange behaviour of the various PrP27-30 samples. FT-IR spectra of PrP27-30 samples from 263K, ME7-H and 22A-H exposed to increasing temperature (up to 90 degrees C) showed that a strain-specific response to heat treatment is associated with strain specific thermostability of distinct secondary structure elements, providing additional means for TSEs strain discrimination.
authors.affiliation: P25, Robert Koch-Institut, Nordufer 20, D-13353 Berlin, Germany.
authors.forename: Sashko
authors.initials: S
authors.lastname: Spassov
authors.forename: Michael
authors.initials: M
authors.lastname: Beekes
authors.forename: Dieter
authors.initials: D
authors.lastname: Naumann
mesh_terms: D000818:Animals; D016000:Cluster Analysis; D006224:Cricetinae; D017666:Deuterium Oxide; D017096:Prion Diseases / Q000378:metabolism*; D011328:Prions / Q000737:chemistry; D011489:Protein Denaturation; D017510:Protein Folding; D017433:Protein Structure, Secondary; D013045:Species Specificity; D013053:Spectrophotometry; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*; D013696:Temperature; D014867:Water
chemical_list: D011328:Prions; D014867:Water; D017666:Deuterium Oxide