Scrapie-infected cells, isolated prions, and recombinant prion protein: a comparative study.
Fourier -transform infrared microscopic spectra of scrapie-infected nervous tissue measured at high spatial resolution (approximately 6 microm) were compared with those obtained from the purified, partly proteinase K digested scrapie isoform of the prion protein isolated from nervous tissue of hamsters infected with the same scrapie strain (263K) to elucidate similarities/dissimilarities between prion structure investigated in situ and ex vivo. A further comparison is drawn to the recombinant Syrian hamster prion protein SHaPrP(90-232) after in vitro conformational transition from the predominantly alpha-helical isoform to beta-sheet-rich structures. It is shown that prion protein structure can be investigated within tissue and that detectability of regions with elevated beta-sheet content as observed in microspectra of prion-infected tissue strongly depends on spatial resolution of the experiment.
Kneipp
J
J
Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany.
Miller
L M
LM
Spassov
S
S
Sokolowski
F
F
Lasch
P
P
Beekes
M
M
Naumann
D
D
D000818:Animals; D006224:Cricetinae; D019286:Endopeptidase K / Q000378:metabolism; D005727:Ganglia, Spinal / Q000378:metabolism; D066298:In Vitro Techniques; D008647:Mesocricetus; D017096:Prion Diseases / Q000378:metabolism; D011328:Prions / Q000737:chemistry*; D011487:Protein Conformation; D020033:Protein Isoforms; D017433:Protein Structure, Secondary; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D012608:Scrapie / Q000378:metabolism*; D017550:Spectroscopy, Fourier Transform Infrared / Q000379:methods*
D011328:Prions; D020033:Protein Isoforms; D011994:Recombinant Proteins; D019286:Endopeptidase K