TY  - JOUR
AU  - Kaimann, T.
AU  - Metzger, S.
AU  - Kuhlmann, K.
AU  - Brandt, B.
AU  - Birkmann, E.
AU  - Höltje, H.-D.
AU  - Riesner, D.
TI  - Molecular Model of an alpha-helical Prion Protein dimer and its monomeric subunits as derived from chemical cross-linking and molecular modeling calculations
JO  - Journal of molecular biology
VL  - 376
SN  - 0022-2836
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - PreJuSER-314
SP  - 582 - 596
PY  - 2008
N1  - Record converted from VDB: 12.11.2012
AB  - Prions are the agents of a series of lethal neurodegenerative diseases. They are composed largely, if not entirely, of the host-encoded prion protein (PrP), which can exist in the cellular isoform PrP(C) and the pathological isoform PrP(Sc). The conformational change of the alpha-helical PrP(C) into beta-sheet-rich PrP(Sc) is the fundamental event of prion disease. The transition of recombinant PrP from a PrP(C)-like into a PrP(Sc)-like conformation can be induced in vitro by submicellar concentrations of SDS. An alpha-helical dimer was identified that might represent either the native state of PrP(C) or the first step from the monomeric PrP(C) to highly aggregated PrP(Sc). In the present study, the molecular structure of these dimers was analyzed by introducing covalent cross-links using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide. Inter- and intramolecular bonds between directly neighboured amino groups and carboxy groups were generated. The bonds formed in PrP dimers of recombinant PrP (90-231) were identified by tryptic digestion and subsequent mass spectrometric analysis. Intra- and intermolecular cross-links between N-terminal glycine and three acidic amino acid side chains in the globular part of PrP were identified, showing the N-terminal amino acids (90-124) are not as flexible as known from NMR analysis. When the cross-linked sites were used as structural constraint, molecular modeling calculations yielded a structural model for PrP dimer and its monomeric subunit, including the folding of amino acids 90-124 in addition to the known structure. Molecular dynamics of the structure after release of the constraint indicated an intrinsic stability of the domain of amino acids 90-124.
KW  - Amino Acids, Acidic: chemistry
KW  - Animals
KW  - Circular Dichroism
KW  - Computer Simulation
KW  - Cricetinae
KW  - Cross-Linking Reagents: chemistry
KW  - Dimerization
KW  - Mesocricetus
KW  - Models, Molecular
KW  - Nuclear Magnetic Resonance, Biomolecular
KW  - Prions: chemistry
KW  - Prions: genetics
KW  - Prions: metabolism
KW  - Protein Conformation
KW  - Protein Isoforms: chemistry
KW  - Protein Isoforms: metabolism
KW  - Protein Structure, Secondary
KW  - Recombinant Proteins: chemistry
KW  - Recombinant Proteins: metabolism
KW  - Spectrometry, Mass, Electrospray Ionization
KW  - Tandem Mass Spectrometry
KW  - Trypsin: pharmacology
KW  - Amino Acids, Acidic (NLM Chemicals)
KW  - Cross-Linking Reagents (NLM Chemicals)
KW  - Prions (NLM Chemicals)
KW  - Protein Isoforms (NLM Chemicals)
KW  - Recombinant Proteins (NLM Chemicals)
KW  - Trypsin (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:18158160
UR  - <Go to ISI:>//WOS:000253554700024
DO  - DOI:10.1016/j.jmb.2007.11.035
UR  - https://juser.fz-juelich.de/record/314
ER  -