Molecular conformation and dynamics of the Y145Stop variant of human prion protein in amyloid fibrils.

@article{citeulike:2742122, abstract = {A C-terminally truncated Y145Stop variant of the human prion protein (huPrP23-144) is associated with a hereditary amyloid disease known as PrP cerebral amyloid angiopathy. Previous studies have shown that recombinant huPrP23-144 can be efficiently converted in vitro to the fibrillar amyloid state, and that residues 138 and 139 play a critical role in the amyloidogenic properties of this protein. Here, we have used magic-angle spinning solid-state NMR spectroscopy to provide high-resolution insight into the protein backbone conformation and dynamics in fibrils formed by (13)C,(15)N-labeled huPrP23-144. Surprisingly, we find that signals from approximately 100 residues (i.e., approximately 80\% of the sequence) are not detected above approximately -20 degrees C in conventional solid-state NMR spectra. Sequential resonance assignments revealed that signals, which are observed, arise exclusively from residues in the region 112-141. These resonances are remarkably narrow, exhibiting average (13)C and (15)N linewidths of approximately 0.6 and 1 ppm, respectively. Altogether, the present findings indicate the existence of a compact, highly ordered core of huPrP23-144 amyloid encompassing residues 112-141. Analysis of (13)C secondary chemical shifts identified likely beta-strand segments within this core region, including beta-strand 130-139 containing critical residues 138 and 139. In contrast to this relatively rigid, beta-sheet-rich amyloid core, the remaining residues in huPrP23-144 amyloid fibrils under physiologically relevant conditions are largely unordered, displaying significant conformational dynamics.}, address = {Department of Chemistry, Ohio State University, Columbus, OH 43210.}, author = {Helmus, J. J. and Surewicz, K. and Nadaud, P. S. and Surewicz, W. K. and Jaroniec, C. P. }, citeulike-article-id = {2742122}, doi = {http://dx.doi.org/10.1073/pnas.0711716105}, issn = {1091-6490}, journal = {Proceedings of the National Academy of Sciences of the United States of America}, keywords = {amyloid, prion}, month = {April}, number = {17}, pages = {6284--6289}, posted-at = {2008-05-01 09:45:13}, priority = {5}, title = {Molecular conformation and dynamics of the Y145Stop variant of human prion protein in amyloid fibrils.}, url = {http://dx.doi.org/10.1073/pnas.0711716105}, volume = {105}, year = {2008} }

TY - JOUR ID - citeulike:2742122 L3 - citeulike-article-id:2742122 TI - Molecular conformation and dynamics of the Y145Stop variant of human prion protein in amyloid fibrils. JF - Proceedings of the National Academy of Sciences of the United States of America VL - 105 IS - 17 SP - 6284 EP - 6289 AD - Department of Chemistry, Ohio State University, Columbus, OH 43210. SN - 1091-6490 N2 - A C-terminally truncated Y145Stop variant of the human prion protein (huPrP23-144) is associated with a hereditary amyloid disease known as PrP cerebral amyloid angiopathy. Previous studies have shown that recombinant huPrP23-144 can be efficiently converted in vitro to the fibrillar amyloid state, and that residues 138 and 139 play a critical role in the amyloidogenic properties of this protein. Here, we have used magic-angle spinning solid-state NMR spectroscopy to provide high-resolution insight into the protein backbone conformation and dynamics in fibrils formed by (13)C,(15)N-labeled huPrP23-144. Surprisingly, we find that signals from approximately 100 residues (i.e., approximately 80% of the sequence) are not detected above approximately -20 degrees C in conventional solid-state NMR spectra. Sequential resonance assignments revealed that signals, which are observed, arise exclusively from residues in the region 112-141. These resonances are remarkably narrow, exhibiting average (13)C and (15)N linewidths of approximately 0.6 and 1 ppm, respectively. Altogether, the present findings indicate the existence of a compact, highly ordered core of huPrP23-144 amyloid encompassing residues 112-141. Analysis of (13)C secondary chemical shifts identified likely beta-strand segments within this core region, including beta-strand 130-139 containing critical residues 138 and 139. In contrast to this relatively rigid, beta-sheet-rich amyloid core, the remaining residues in huPrP23-144 amyloid fibrils under physiologically relevant conditions are largely unordered, displaying significant conformational dynamics. KW - amyloid KW - prion AU - Helmus, JJ AU - Surewicz, K AU - Nadaud, PS AU - Surewicz, WK AU - Jaroniec, CP PY - 2008/04/29/ UR - http://dx.doi.org/10.1073/pnas.0711716105 ER -