Designing conditions for <i>in vitro</i> formation of amyloid protofilaments and fibrils

• Fabrizio Chiti

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Paul Webster

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Niccolò Taddei

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Anne Clark

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Massimo Stefani

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Giampietro Ramponi

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

• Christopher M. Dobson

  Oxford Centre for Molecular Sciences, New Chemistry Laboratory and Department of Human Anatomy and Genetics, University of Oxford, South Parks Road, Oxford OX1 3QT, United Kingdom; and Dipartimento di Scienze Biochimiche, Università degli Studi di Firenze, Viale Morgagni 50, 50134 Firenze, Italy

抄録

We have been able to convert a small α/β protein, acylphosphatase, from its soluble and native form into insoluble amyloid fibrils of the type observed in a range of pathological conditions. This was achieved by allowing slow growth in a solution containing moderate concentrations of trifluoroethanol. When analyzed with electron microscopy, the protein aggregate present in the sample after long incubation times consisted of extended, unbranched filaments of 30–50 Å in width that assemble subsequently into higher order structures. This fibrillar material possesses extensive β-sheet structure as revealed by far-UV CD and IR spectroscopy. Furthermore, the fibrils exhibit Congo red birefringence, increased fluorescence with thioflavine T and cause a red-shift of the Congo red absorption spectrum. All of these characteristics are typical of amyloid fibrils. The results indicate that formation of amyloid occurs when the native fold of a protein is destabilized under conditions in which noncovalent interactions, and in particular hydrogen bonding, within the polypeptide chain remain favorable. We suggest that amyloid formation is not restricted to a small number of protein sequences but is a property common to many, if not all, natural polypeptide chains under appropriate conditions.

収録刊行物

• Proceedings of the National Academy of Sciences

  Proceedings of the National Academy of Sciences 96 (7), 3590-3594, 1999-03-30

  Proceedings of the National Academy of Sciences

キーワード

• Multidisciplinary

詳細情報

• CRID: 1360855570273565824
• NII論文ID: 80011067582
• DOI: 10.1073/pnas.96.7.3590
• ISSN: 10916490
• ISSN: 00278424
• Web Site: https://pnas.org/doi/pdf/10.1073/pnas.96.7.3590
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