Familial Mutations and the Thermodynamic Stability of the Recombinant Human Prion Protein

• Published in: The Journal of biological chemistry Vol. 273; no. 47; pp. 31048 - 31052
• Main Authors: Swietnicki, Wieslaw, Petersen, Robert B., Gambetti, Pierluigi, Surewicz, Witold K.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 20.11.1998; American Society for Biochemistry and Molecular Biology
• Subjects: Creutzfeldt-Jakob Syndrome - genetics; Gerstmann-Straussler-Scheinker Disease - genetics; Guanidine; Hot Temperature; Humans; Peptide Fragments - chemistry; Peptide Fragments - genetics; Point Mutation; Prion Diseases - genetics; Prions - chemistry; Prions - genetics; Protein Denaturation; Recombinant Proteins - chemistry; Spectrometry, Fluorescence; Thermodynamics
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1074/jbc.273.47.31048

Hereditary forms of human prion disease are linked to specific mutations in the PRNP gene. It has been postulated that these mutations may facilitate the pathogenic process by reducing the stability of the prion protein (PrP). To test this hypothesis, we characterized the recombinant variants of human PrP(90–231) containing point mutations corresponding to Gerstmann-Straussler-Scheinker disease (P102L), Creutzfeld-Jakob disease (E200K), and fatal familial insomnia (M129/D178N). The first two of these mutants could be recovered form from the periplasmic space of Escherichia coli in a soluble form, whereas the D178N variant aggregated into inclusion bodies. The secondary structure of the two soluble variants was essentially identical to that of the wild-type protein. The thermodynamic stability of these mutants was assessed by unfolding in guanidine hydrochloride and thermal denaturation. The stability properties of the P102L variant were indistinguishable from those of wild-type PrP, whereas the E200K mutation resulted in a very small destabilization of the protein. These data, together with the predictive analysis of other familial mutations, indicate that some hereditary forms of prion disease cannot be rationalized using the concept of mutation-induced thermodynamic destabilization of the cellular prion protein.