Amyloid fibril formation by a partially structured intermediate state of α-chymotrypsin

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Here we investigated the effects of 2,2,2-trifluoroethanol (TFE) on the structure of α-chymotrypsin. The protein aggregates maximally in 35% (v/v) TFE. Congo red and thioflavin-T binding experiments suggest that the aggregates induced by TFE have amyloid-like properties, and transmission electron microscopy data show that these aggregates have a fibrilar morphology. Fluorescence, circular dichroism, anilino-8-napthalene sulfonate binding, and Fourier-transformed infrared spectroscopy data suggest that formation of a partially structured intermediate state precedes the onset of the aggregation process. The native β-barrel structure of α-chymotrypsin appears to be disrupted in the partially structured intermediate state in favour of a non-native extended β-sheet conformation with exposed hydrophobic surfaces. The protein becomes "sticky" under these conditions and aggregates into amyloid-like structures. The data support the hypothesis that amyloid formation involves the ordered self-assembly of partially folded species that are critical soluble precursors of fibrilar aggregates. © 2004 Elsevier Ltd. All rights reserved.
Original languageEnglish
Pages (from-to)321-331
JournalJournal of Molecular Biology
Issue number1
Publication statusPublished - 3 Sep 2004


  • α-chymotrypsin
  • aggregation intermediates
  • amyloid formation
  • protein misfolding
  • serine proteases
  • TFE-induced protein denaturation


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