title: Unraveling the Molecular Mechanism of pH-Induced Misfolding and Oligomerization of the Prion Protein.
abstract: The misfolding of the prion protein (PrP) to aggregated forms is linked to several neurodegenerative diseases. Misfolded oligomeric forms of PrP are associated with neurotoxicity and/or infectivity, but the molecular mechanism by which they form is still poorly understood. A reduction in pH is known to be a key factor that triggers misfolded oligomer formation by PrP, but the residues whose protonation is linked with misfolding remain unidentified. The structural consequences of the protonation of these residues also remain to be determined. In the current study, amino acid residues whose protonation is critical for PrP misfolding and oligomerization have been identified using site-directed mutagenesis and misfolding/oligomerization assays. It is shown that the protonation of either H186 or D201, which mimics the effects of pathogenic mutations (H186R and D201N) at both residue sites, is critically linked to the stability, misfolding and oligomerization of PrP. Hydrogen-deuterium exchange studies coupled with mass spectrometry show that the protonation of either H186 or D201 leads to the same common structural change: increased structural dynamics in helix 1 and that in the loop between helix 1 and β-strand 2. It is shown that the protonation of either of these residues is sufficient for accelerating misfolded oligomer formation, most likely because the protonation of either residue causes the same structural perturbation. Hence, the increased structural dynamics in helix 1 and that in the loop between helix 1 and β-strand 2 appear to play an early critical role in acid-induced misfolding of PrP.
authors: National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru 560065, India.
authors: Jogender
authors: J
authors: Singh
authors: National Centre for Biological Sciences, Tata Institute of Fundamental Research, Bengaluru 560065, India. Electronic address: jayant@ncbs.res.in.
authors: Jayant B
authors: JB
authors: Udgaonkar
mesh_terms: D004252:DNA Mutational Analysis; D006863:Hydrogen-Ion Concentration; D013058:Mass Spectrometry; D016297:Mutagenesis, Site-Directed; D011328:Prions / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D066263:Protein Aggregation, Pathological; D017510:Protein Folding / Q000187:drug effects*; D055503:Protein Multimerization; D055550:Protein Stability
chemical_list: D011328:Prions
keywords: D201N; H186R; hydrogen exchange; misfolding; prion protein