NR ARQG
AU Hornemann,S.; Schorn,C.; Wüthrich,K.
TI NMR structure of the bovine prion protein isolated from healthy calf brains
QU EMBO Reports 2004 Dec; 5(12): 1159-64
IA http://www.nature.com/embor/journal/v5/n12/pdf/7400297.pdf
PT journal article
AB NMR structures of recombinant prion proteins from various species expressed in Escherichia coli have been solved during the past years, but the fundamental question of the relevancy of these data relative to the naturally occurring forms of the prion protein has not been directly addressed. Here, we present a comparison of the cellular form of the bovine prion protein isolated and purified from healthy calf brains without use of detergents, so that it contains the two carbohydrate moieties and the part of the GPI anchor that is maintained after enzymatic cleavage of the glycerolipid moiety, with the recombinant bovine prion protein expressed in E. coli. We show by circular dichroism and (1)H-NMR spectroscopy that the three-dimensional structure and the thermal stability of the natural glycoprotein and the recombinant polypeptide are essentially identical. This result indicates possible functional roles of the glycosylation of prion proteins in healthy organisms, and provides a platform and validation for future work on the structural biology of prion proteins, which will have to rely primarily on the use of recombinant polypeptides.
MH Animals; Brain/*metabolism; Cattle; Circular Dichroism; Magnetic Resonance Spectroscopy; Prions/*chemistry/isolation & purification; Protein Structure, Secondary; Protein Structure, Tertiary; Research Support, Non-U.S. Gov't; Temperature
AD Institut für Molekularbiologie und Biophysik, Eidgenössische Technische Hochschule Zürich, 8093 Zürich, Switzerland. hornemann@mol.biol.ethz.ch
SP englisch
PO England