The mammalian prion protein, PrP^C, is able to switch its conformation from a monomeric soluble form into an aggregated transmissible prion state, PrP^Sc. Once a prion is introduced into a susceptible host it triggers a PrP conversion cascade, which leads to prion disease. However, a molecular understanding of the involved processes and determinants is missing. Multiple studies have converted recombinant PrP into amyloid-like forms, but whose relation to prion disease needs further investigation.

In this project, recombinant PrP will be produced in E. coli using an established purification protocol. The monomeric recombinant PrP will then be converted de novo into fibrillar aggregates using published procedures. The sequence specific location of regular secondary structures of the aggregates will then be investigated by quenched hydrogen exchange NMR (qHDX-NMR). During the project, suitable experimental conditions for qHDX-NMR have to be established, and the NMR chemical shift assignment of the backbone amides has to be determined. The knowledge of the sequence specific location of regular secondary structures will be a crucial step towards an understanding the molecular mechanism underlying the conformational conversion of PrP.

The successfull candidate will be introduced into all nescessary molecular biological and protein biochemical techniques. The obtained products will be first characterized by multiple biophysical techniques including NMR-spectroscopy, CD-spectroscopy, fourier transformed infrared spectroscopy and light scattering. The project will be carried out in close collaboration with other ongoing projects in the laboratory.