Structural changes in a hydrophobic domain of the prion protein induced by hydration and by Ala → Val and Pro → Leu substitutions

X-ray diffraction was used to study the structure of assemblies formed by synthetic peptide fragments of the prion protein (PrP) that include the hydrophobic domain implicated in the Gerstmann-Straussler-Scheinker (GSS) mutation (P102L). The effects of hydration on polypeptide assembly and of Ala → Val substitutions in the hydrophobic domain were characterized. Synthetic peptides included: (i) Syrian hamster (SHa) hydrophobic core, SHa106-122 (KTNMKHMAGAAAAGAVV); (ii) SHa104-122(3A-V), with A → V mutations at 113, 115 and 118 (KPKTNMKHMVGVAAVGAVV); (iii) mouse (Mo) wild-type sequence of the N-terminal hydrophobic domain, Mo89-143WT; and (iv) the same mouse sequence with leucine substitution for proline at residue number 101, Mo89-143(P101L). Samples of SHa106-122 that formed assemblies while drying tinder ambient conditions showed X-ray patterns indicative of 33 Å thick slab-like structures having extensive H-bonding and intersheet stacking. By contrast, lyophilized peptide that was equilibrated against 100% relative humidity showed assemblies with only a few layers of β-sheets. The Ala → Val substitutions in SHa104-122 and Mo89-143(P101L) resulted in the formation of 40 Å wide, cross-β fibrils. Observation of similar size β-sheet fibrils formed by peptides SHa104-122(3A-V) and the longer Mo89-143(P101L) supports the notion that the hydrophobic sequence forms a template or core that promotes the β-folding of the longer peptide. The substitution of amino acids in the mutants, e.g. 3A → V and P101L, enhances the folding of the peptide into compact structural units, significantly enhancing the formation of the extensive β-sheet fibrils. (C) 2000 Academic Press.