| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
1 Medical Biotechnology Center, University of Maryland Biotechnology Institute, Baltimore, Maryland 21201, USA2 Department of Biochemistry and Molecular Biology, University of Maryland School of Medicine, Baltimore, Maryland 21201, USA3 Institute for Animal Health, Compton, Newbury, Berkshire, RG20 7NN, United Kingdom
(RECEIVED October 20, 2004; FINAL REVISION January 17, 2005; ACCEPTED January 23, 2005)
In recent studies, the amyloid form of recombinant prion protein (PrP) encompassing residues 89–230 (rPrP 89-230) produced in vitro induced transmissible prion disease in mice. These studies showed that unlike "classical" PrPSc produced in vivo, the amyloid fibrils generated in vitro were more proteinase-K sensitive. Here we demonstrate that the amyloid form contains a proteinase K-resistant core composed only of residues 152/153–230 and 162–230. The PK-resistant fragments of the amyloid form are similar to those observed upon PK digestion of a minor subpopulation of PrPSc recently identified in patients with sporadic Creutzfeldt-Jakob disease (CJD). Remarkably, this core is sufficient for self-propagating activity in vitro and preserves a 
-sheet-rich fibrillar structure. Full-length recombinant PrP 23-230, however, generates two subpopulations of amyloid in vitro: One is similar to the minor subpopulation of PrPSc, and the other to classical PrPSc. Since no cellular factors or templates were used for generation of the amyloid fibrils in vitro, we speculate that formation of the subpopulation of PrPSc with a short PK-resistant C-terminal region reflects an intrinsic property of PrP rather than the influence of cellular environments and/or cofactors. Our work significantly increases our understanding of the biochemical nature of prion infectious agents and provides a fundamental insight into the mechanisms of prions biogenesis.
Keywords: prion protein; amyloid fibrils; conformational transition; proteinase K; Creutzfeldt-Jakob disease
Abbreviations: CJD, Creutzfeldt-Jakob disease • spCJD, sporadic CJD • PrP, prion protein • PrPC, the normal, cellular isoform of PrP • PrPSc, the abnormal, infections isoform of PrP • rPrP, recombinant PrP • rPrP 89-230, recombinant PrP encompassing residues 89–230 • rPrP 23-230, full-length recombinant PrP • 
-rPrP 89-230, -helical isoform of rPrP 89-231 • -rPrP 23-230, -helical isoform of rPrP 23-231 • PK, proteinase K • PrP 27-30, PK-resistant core of classical PrPSc • ThT, Thioflavin T • GdnHCl, guanidinium hydrochloride • FTIR; Fourier transform infrared spectroscopy
Article and publication are at http://www.proteinscience.org/cgi/doi/10.1110/ps.041186605.
Reprint requests to: Ilia V. Baskakov, 725 W. Lombard Street, Baltimore, MD 21201, USA; e-mail: Baskakov{at}umbi.umd.edu; fax: (410) 706-8184.
CiteULike 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  N. Makarava and I. V. Baskakov The Same Primary Structure of the Prion Protein Yields Two Distinct Self-propagating States J. Biol. Chem., June 6, 2008; 283(23): 15988 - 15996. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. J. Helmus, K. Surewicz, P. S. Nadaud, W. K. Surewicz, and C. P. Jaroniec Molecular conformation and dynamics of the Y145Stop variant of human prion protein in amyloid fibrils PNAS, April 29, 2008; 105(17): 6284 - 6289. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. Stohr, N. Weinmann, H. Wille, T. Kaimann, L. Nagel-Steger, E. Birkmann, G. Panza, S. B. Prusiner, M. Eigen, and D. Riesner Mechanisms of prion protein assembly into amyloid PNAS, February 19, 2008; 105(7): 2409 - 2414. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
N. Makarava, C.-I Lee, V. G. Ostapchenko, and I. V. Baskakov Highly Promiscuous Nature of Prion Polymerization J. Biol. Chem., December 14, 2007; 282(50): 36704 - 36713. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 C. I. Lee, Q. Yang, V. Perrier, and I. V. Baskakov The dominant-negative effect of the Q218K variant of the prion protein does not require protein X Protein Sci., October 1, 2007; 16(10): 2166 - 2173. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Sun, L. Breydo, N. Makarava, Q. Yang, O. V. Bocharova, and I. V. Baskakov Site-specific Conformational Studies of Prion Protein (PrP) Amyloid Fibrils Revealed Two Cooperative Folding Domains within Amyloid Structure J. Biol. Chem., March 23, 2007; 282(12): 9090 - 9097. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
X. Lu, P. L. Wintrode, and W. K. Surewicz beta-Sheet core of human prion protein amyloid fibrils as determined by hydrogen/deuterium exchange PNAS, January 30, 2007; 104(5): 1510 - 1515. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. M. Martins, D. J. Frosoni, A. M. B. Martinez, F. G. De Felice, and S. T. Ferreira Formation of Soluble Oligomers and Amyloid Fibrils with Physical Properties of the Scrapie Isoform of the Prion Protein from the C-terminal Domain of Recombinant Murine Prion Protein mPrP-(121-231) J. Biol. Chem., September 8, 2006; 281(36): 26121 - 26128. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Novitskaya, N. Makarava, A. Bellon, O. V. Bocharova, I. B. Bronstein, R. A. Williamson, and I. V. Baskakov Probing the Conformation of the Prion Protein within a Single Amyloid Fibril Using a Novel Immunoconformational Assay J. Biol. Chem., June 2, 2006; 281(22): 15536 - 15545. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
O. V. Bocharova, N. Makarava, L. Breydo, M. Anderson, V. V. Salnikov, and I. V. Baskakov Annealing Prion Protein Amyloid Fibrils at High Temperature Results in Extension of a Proteinase K-resistant Core J. Biol. Chem., January 27, 2006; 281(4): 2373 - 2379. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
