The structural intolerance of the PrP alpha-fold for polar substitution of the helix-3 methionines.
The conversion of the cellular prion protein (PrP(C)) into its disease-associated form (PrP(Sc)) involves a major conformational change and the accumulation of sulfoxidized methionines. Computational and synthetic approaches have shown that this change in the polarity of M206 and M213 impacts the C-terminal domain native alpha-fold allowing the flexibility required for the structural conversion. To test the effect in the full-length molecule with site-specificity, we have generated M-to-S mutations. Molecular dynamics simulations show that the replacement indeed perturbs the native state. When this mutation is placed at the conserved methionines of HaPrP(23-231), only substitutions at the Helix-3 impair the alpha-fold, stabilizing a non-native state with perturbed secondary structure, loss of native tertiary contacts, increased surface hydrophobicity, reduced thermal stability and an enhanced tendency to aggregate into protofibrillar polymers. Our work supports that M206 and M213 function as alpha-fold gatekeepers and suggests that their redox state regulate misfolding routes.
Lisa
Silvia
S
Insto Química-Física Rocasolano, CSIC, 28006 Madrid, Spain.
Meli
Massimiliano
M
Cabello
Gema
G
Gabizon
Ruth
R
Colombo
Giorgio
G
Gasset
María
M
D019943:Amino Acid Substitution; D002942:Circular Dichroism; D017931:DNA Primers; D014644:Genetic Variation; D006714:Homoserine / Q000031:analogs & derivatives / Q000378:metabolism; D006801:Humans; D008715:Methionine / Q000378:metabolism*; D008958:Models, Molecular; D009646:Norleucine / Q000378:metabolism; D017096:Prion Diseases / Q000235:genetics / Q000378:metabolism*; D011328:Prions / Q000737:chemistry* / Q000235:genetics; D011487:Protein Conformation; D017510:Protein Folding
D017931:DNA Primers; D011328:Prions; C037855:methoxine; D006714:Homoserine; D009646:Norleucine; D008715:Methionine
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