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Seeded conversion of recombinant prion protein to a disulfide-bonded oligomer by a reduction-oxidation process

Nature Structural & Molecular Biology volume 10, pages 725–730 (2003)Cite this article

Abstract

The infectious form of prion protein, PrPSc, self-propagates by its conversion of the normal, cellular prion protein molecule PrPC to another PrPSc molecule. It has not yet been demonstrated that recombinant prion protein can convert prion protein molecules from PrPC to PrPSc. Here we show that recombinant hamster prion protein is converted to a second form, PrPRDX, by a redox process in vitro and that this PrPRDX form seeds the conversion of other PrPC molecules to the PrPRDX form. The converted form shows properties of oligomerization and seeded conversion that are characteristic of PrPSc. We also find that the oligomerization can be reversed in vitro. X-ray fiber diffraction suggests an amyloid-like structure for the oligomerized prion protein. A domain-swapping model involving intermolecular disulfide bonds can account for the stability and coexistence of two molecular forms of prion protein and the capacity of the second form for self-propagation.

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Figure 1: Conversion of the monomeric HisPrP(90–231) to an oligomeric form by a redox process.
Figure 2: Conversion of PrPC to PrPRDX by intermolecular disulfide linkage.
Figure 3: Seeded oligomerization of HisPrP(90–231) by the redox process.
Figure 4: Partial proteinase K (PK) resistance of HisPrPRDX(23–231).
Figure 5: Structural features of HisPrPRDX(90–231).
Figure 6: Speculative model for conversion of PrPC (lower right) to a PrPRDX fibril (lower left).

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Acknowledgements

We thank S.B. Prusiner's laboratory for helping us in fermentation of HisPrP(90–231), M. Gingery for electron microscopy, M.R. Sawaya for building the model of the PrPRDX fibril, R.L. Garrell's laboratory for helping us in g-factor analysis, M. Apostol for assistance and P.D. Boyer, T.E. Creighton and A.K. Chamberlain for critical reading of the manuscript. This work was supported by the Howard Hughes Medical Institute and the US National Institutes of Health.

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  1. UCLA-DOE Institute for Genomics and Proteomics and Department of Chemistry and Biochemistry, Howard Hughes Medical Institute, Molecular Biology Institute, University of California, Los Angeles, Los Angeles, 90095-1570, California, USA

    Sangho Lee & David Eisenberg

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Correspondence to David Eisenberg.

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Lee, S., Eisenberg, D. Seeded conversion of recombinant prion protein to a disulfide-bonded oligomer by a reduction-oxidation process. Nat Struct Mol Biol 10, 725–730 (2003). https://doi.org/10.1038/nsb961

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