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

Hideyo Inouye, Jeremy Bond, Michael A. Baldwin, Haydn L. Ball, Stanley B. Prusiner, Daniel A. Kirschner

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)1283-1296
Number of pages14
JournalJournal of Molecular Biology
Volume300
Issue number5
DOIs
StatePublished - Jul 28 2000

Fingerprint

Peptides
Mutation
Peptide Fragments
Mesocricetus
Amino Acid Substitution
Humidity
Proline
Leucine
X-Ray Diffraction
Prion Proteins
Observation
X-Rays

Keywords

  • β-pleated sheet
  • Amyloidogenic peptide
  • Gerstmann-Straussler-Scheinker
  • Prion
  • X-ray diffraction

ASJC Scopus subject areas

  • Virology

Cite this

Structural changes in a hydrophobic domain of the prion protein induced by hydration and by Ala → Val and Pro → Leu substitutions. / Inouye, Hideyo; Bond, Jeremy; Baldwin, Michael A.; Ball, Haydn L.; Prusiner, Stanley B.; Kirschner, Daniel A.

In: Journal of Molecular Biology, Vol. 300, No. 5, 28.07.2000, p. 1283-1296.

Research output: Contribution to journalArticle

Inouye, Hideyo ; Bond, Jeremy ; Baldwin, Michael A. ; Ball, Haydn L. ; Prusiner, Stanley B. ; Kirschner, Daniel A. / Structural changes in a hydrophobic domain of the prion protein induced by hydration and by Ala → Val and Pro → Leu substitutions. In: Journal of Molecular Biology. 2000 ; Vol. 300, No. 5. pp. 1283-1296.
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abstract = "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 {\AA} 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 {\AA} 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.",
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T1 - Structural changes in a hydrophobic domain of the prion protein induced by hydration and by Ala → Val and Pro → Leu substitutions

AU - Inouye, Hideyo

AU - Bond, Jeremy

AU - Baldwin, Michael A.

AU - Ball, Haydn L.

AU - Prusiner, Stanley B.

AU - Kirschner, Daniel A.

PY - 2000/7/28

Y1 - 2000/7/28

N2 - 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.

AB - 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.

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