NR ATRQ
AU Sasaki,K.; Gaikwad,J.U.; Akasaka,K.; Kitamoto,S.; Hashiguchi,S.; Sugimura,K.
TI Characterization of oligomeric forms of human Prion Protein HuPrPc (23-231) by CD, AFM, and Fluorescence spectroscopy.
QU International Conference - Prion 2005: Between fundamentals and society's needs - 19.10.-21.10.2005, Congress Center Düsseldorf - Poster Session: Structure of PrP and molecular determinants of infectivity STRCT-32
PT Konferenz-Poster
AB
Monomer to oligomer transition is a key to understanding the molecular mechanism of the transformation of PrPc to PrPsc. Recently, the method of preparation of b-sheet-rich forms of PrPc has been reported for both intact and reduced Human Prion Protein HuPrPc (90-231) [1,2]. In the present work, we have similarly prepared the b-rich forms of HuPrPc (23-231) with and without the reduced disulfide bond and tried to characterize their structures and stability by using CD, fluorescence and Atomic Force Microscopy. The CD spectra clearly showed that the original alpha-rich structure of HuPrPc (23-231) had turned into a beta-rich structure. The AFM images showed many globular shaped spots, which suggested the formation of oligomeric HuPrPc (23-231) without forming fibrils. The solution containing the oligomeric HuPrPc (23-231) showed strong fluorescence from eight Trp residues, which upon dilution, sharply decreased with time. After the dilution, the globular shaped spots in AFM have disappeared, suggesting the dissociation of the oligomers into monomeric species.
[1] Reversible Conversion of Monomeric Human Prion Protein Between Native and Fibrilogenic Conformations, G. S. Jackson, L. L. P. Hosszu, A. Power, A. F. Hill, J. Kenney, H. Saibil, C. J. Craven, J. P. Waltho, A. R. Clarke, and J. Collinge, Science 283: 1935-1937. 1999
[2] Pathway Complexity of Prion Protein Assembly into Amyloid, I.V. Baskakov, G. Legname, M.A. Baldwin, S.B. Pruciner, F.E. Cohen, J. Biol. Chem. 277: 21140-21148, 2002
AD K.Sasaki, J.U.Gaikwad, K.Akasaka, Department of Biotechnological Science, School of Biology-Oriented Science & Technology, Japan; S.Kitamoto, S.Hashiguchi, K.Sugimura, Department of Bioengineering, Faculty of Engineering, Kagoshima University
SP englisch
PO Deutschland