NR AFON
AU Hornemann,S.; Korth,C.; Oesch,B.; Riek,R.; Wider,G.; Wüthrich,K.; Glockshuber,R.
TI Recombinant full-length murine prion protein, mPrP(23-231): purification and spectroscopic characterization.
QU FEBS Letters 1997 Aug 18; 413(2): 277-81
PT journal article
AB The cellular prion protein of the mouse, mPrPc, consists of 208 amino acids (residues 23-231). It contains a carboxy-terminal domain, mPrP(121-231), which represents an autonomous folding unit with three alpha-helices and a two-stranded antiparallel beta-sheet. We expressed the complete amino acid sequence of the prion protein, mPrP(23-231), in the cytoplasm of Escherichia coli. mPrP(23-231) was solubilized from inclusion bodies by 8 M urea, oxidatively refolded and purified to homogeneity by conventional chromatographic techniques. Comparison of near-UV circular dichroism, fluorescence and one-dimensional 1H-NMR spectra of mPrP(23-231) and mPrP(121-231) shows that the amino-terminal segment 23-120, which includes the five characteristic octapeptide repeats, does not contribute measurably to the manifestation of three-dimensional structure as detected by these techniques, indicating that the residues 121-231 might be the only polypeptide segment of PrPc with a defined three-dimensional structure.
ZR 27
MH Amino Acid Sequence; Animal; Base Sequence; Chromatography, Ion Exchange; Circular Dichroism; Disulfides/chemistry; Magnetic Resonance Spectroscopy; Mice; Molecular Sequence Data; Oxidation-Reduction; Peptide Fragments/chemistry; PrPc Proteins/*chemistry/genetics/*isolation & purification; Protein Folding; Protein Structure, Secondary; Protein Structure, Tertiary; Recombinant Fusion Proteins/chemistry/isolation & purification; Spectrometry, Fluorescence; Support, Non-U.S. Gov't
AD Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Hönggerberg, Zürich, Switzerland.
SP englisch
PO Niederlande
OR Prion-Krankheiten H