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Journal Articles Journal of Molecular Biology Year : 2001

Dependence on solution conditions of aggregation and amyloid formation by an SH3 domain.

Abstract

The formation of amyloid fibrils by the SH3 domain of the alpha-subunit of bovine phosphatidylinositol-3'-kinase (PI3-SH3) has been investigated under carefully controlled solution conditions. NMR and CD characterisation of the denatured states from which fibrils form at low pH show that their properties can be correlated with the nature of the resulting aggregates defined by EM and FTIR spectroscopy. Compact partially folded states, favoured by the addition of anions, are prone to precipitate rapidly into amorphous species, whilst well-defined fibrillar structures are formed slowly from more expanded denatured states. Kinetic data obtained by a variety of techniques show a clear lag phase in the formation of amyloid fibrils. NMR spectroscopy shows no evidence for a significant population of small oligomers in solution during or after this lag phase. EM and FTIR indicate the presence of amorphous aggregates (protofibrils) rich in beta-structure after the lag phase but prior to the development of well-defined amyloid fibrils. These observations strongly suggest a nucleation and growth mechanism for the formation of the ordered aggregates. The morphologies of the fibrillar structures were found to be highly sensitive to the pH at which the protein solutions are incubated. This can be attributed to the effect of small perturbations in the electrostatic interactions that stabilise the contacts between the protofilaments forming the amyloid fibrils. Moreover, different hydrogen bonding patterns related to the various aggregate morphologies can be distinguished by FTIR analysis.
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Dates and versions

pasteur-00364873 , version 1 (27-02-2009)

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J. Zurdo, J. I. Guijarro, J. L. Jiménez, H. R. Saibil, C. M. Dobson. Dependence on solution conditions of aggregation and amyloid formation by an SH3 domain.. Journal of Molecular Biology, 2001, 311 (2), pp.325-40. ⟨10.1006/jmbi.2001.4858⟩. ⟨pasteur-00364873⟩

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