Recombinant full-length murine prion protein, mPrP(23-231):
purification and spectroscopic characterization.

Hornemann S, Korth C, Oesch B, Riek R, Wider G, Wuthrich K, Glockshuber R

Institut fur Molekularbiologie und Biophysik, Eidgenossische Technische Hochschule Honggerberg,
Zurich, Switzerland.

The cellular prion protein of the mouse, mPrP(C), 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 PrP(C) with
a defined three-dimensional structure. 

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