In this application we propose to continue our studies aimed on understanding the molecular basis of the pathogenic process in transmissible spongiform encephalopathies, a group of fatal neurodegenerative diseases such as Creutzfeldt-Jakob disease in humans and bovine spongiform encephalopathy in cattle. It is believed that the infectious pathogen responsible for these diseases is a misfolded, proteinase-resistant form of the prion protein, PrPSc, which is derived from a precursor PrPC by a poorly understood mechanism. While the structure of PrPC is well characterized, one of the most enduring mysteries in the field is the molecular structure of infectious PrPSc conformers. In this project, we propose to undertake a major effort to bridge the gap in understanding the structural properties of abnormally folded prion protein aggregates, with specific aims focused on: (i) determining the conformational structure of proteinase K- resistant form of the recombinant Syrian hamster prion protein, PrPres, obtained by brain PrPSc- templated protein misfolding cyclic amplification (PMCA) reaction;(ii) determining medium- resolution structure (site- and region-specific structural constraints) for brain-derived hamster PrPSc;(iii) elucidating the nature of structural differences between PrPSc corresponding to the hyper and drowsy strains of TSE. This study will employ a number of complementary biophysical methods including site-directed spin labeling, solid-state NMR spectroscopy, hydrogen/deuterium exchange, and hydroxyl radical-mediated footprinting coupled with mass spectrometry.
Prions are infectious proteins that are believed to cause transmissible spongiform encephalopathies (TSEs), a group of fatal neurological disorders including Creutzfeldt-Jakob disease in humans, `mad cow'disease in cattle, and chronic wasting disease in cervids. Understanding the molecular basis of these disorders and the mechanism of their transmissibility is of major importance for public health, especially in view of indications that some animal forms of TSE diseases may cross interspecies transmissibility barriers, posing a threat to humans.
|Nieznanski, Krzysztof; Surewicz, Krystyna; Chen, Shugui et al. (2014) Interaction between prion protein and A? amyloid fibrils revisited. ACS Chem Neurosci 5:340-5|
|Cobb, Nathan J; Apostol, Marcin I; Chen, Shugui et al. (2014) Conformational stability of mammalian prion protein amyloid fibrils is dictated by a packing polymorphism within the core region. J Biol Chem 289:2643-50|
|Apostol, Marcin I; Perry, Kay; Surewicz, Witold K (2013) Crystal structure of a human prion protein fragment reveals a motif for oligomer formation. J Am Chem Soc 135:10202-5|
|Dutta, Arpana; Chen, Shugui; Surewicz, Witold K (2013) The effect of ?2-?2 loop mutation on amyloidogenic properties of the prion protein. FEBS Lett 587:2918-23|
|Kong, Qingzhong; Mills, Jeffrey L; Kundu, Bishwajit et al. (2013) Thermodynamic stabilization of the folded domain of prion protein inhibits prion infection in vivo. Cell Rep 4:248-54|
|Nieznanski, Krzysztof; Choi, Jin-Kyu; Chen, Shugui et al. (2012) Soluble prion protein inhibits amyloid-? (A?) fibrillization and toxicity. J Biol Chem 287:33104-8|
|Helmus, Jonathan J; Surewicz, Krystyna; Apostol, Marcin I et al. (2011) Intermolecular alignment in Y145Stop human prion protein amyloid fibrils probed by solid-state NMR spectroscopy. J Am Chem Soc 133:13934-7|
|Surewicz, Witold K; Apostol, Marcin I (2011) Prion protein and its conformational conversion: a structural perspective. Top Curr Chem 305:135-67|
|Smirnovas, Vytautas; Baron, Gerald S; Offerdahl, Danielle K et al. (2011) Structural organization of brain-derived mammalian prions examined by hydrogen-deuterium exchange. Nat Struct Mol Biol 18:504-6|
|Jones, Eric M; Wu, Bo; Surewicz, Krystyna et al. (2011) Structural polymorphism in amyloids: new insights from studies with Y145Stop prion protein fibrils. J Biol Chem 286:42777-84|
Showing the most recent 10 out of 26 publications