Folding kinetics of the human prion protein probed by temperature jump.

Tanya Hart, Laszlo L P Hosszu, Clare R Trevitt, Graham S Jackson, Jonathan P Waltho, John Collinge, Anthony R Clarke

    Research output: Contribution to journalArticlepeer-review


    Temperature-jump perturbation was used to examine the relaxation kinetics of folding of the human prion protein. Measured rates were very fast (approximately 3,000 s(-1)), with the extrapolated folding rate constant at approximately 20 degrees C in physiological conditions reaching 20,000 s(-1). By a mutational analysis of core residues, we found that only 2, on the interface of helices 2 and 3, have significant phi-values in the transition state. Interestingly, a mutation sandwiched between the above 2 residues on the helix-helix contact interface had very little effect on the overall free energy of folding but led to the formation of a monomeric misfolded state, which had to unfold to acquire the native PrP(C) conformation. Another mutation that led to a marked destabilization of the native fold also formed a misfolded intermediate, but this was aggregation-prone despite the native state of this mutant being soluble. Taken together, the data imply that this fast-folding protein has a transition state that is not compact (m value analysis gives a beta(t) value of only 0.3) but contains a developing nucleus between helices 2 and 3. The fact that a mutation in this nucleus had a negligible effect on stability but still led to formation of aberrant conformations during folding implies an easily perturbed folding mechanism. It is notable that in inherited forms of human prion disease, where point mutations produce a lethal dominant condition, 20 of the 33 amino acid replacements occur in the helix-2/3 sequence.
    Original languageEnglish
    Pages (from-to)5651-5656
    Number of pages5
    JournalProceedings of the National Academy of Sciences of the United States of America
    Issue number14
    Publication statusPublished - 7 Apr 2009


    • φ-value analysis
    • Equilibrium perturbation
    • Protein folding


    Dive into the research topics of 'Folding kinetics of the human prion protein probed by temperature jump.'. Together they form a unique fingerprint.

    Cite this