amyloid fibril formation by bovine cytochrome c淀粉样原纤维形成的牛细胞色素c.pdfVIP

Spectroscopy 19 (2005) 199–205 199 IOS Press Amyloid fibril formation by bovine cytochrome c Natalia S. de Groot and Salvador Ventura ∗ Departament de Bioquimica i Biologia Molecular, Universitat Autonoma de Barcelona and Institut de Biotecnologia i de Biomedicina, 08193 Bellaterra (Barcelona), Spain Abstract. Bovine heart cytochrome c is an all-α globular protein containing a covalently bound heme group. Prolonged incu- bation at 75◦C in mild alkaline solution damages the prosthetic group and results in permanent unfolding of the polypeptide chain. Under this conditions, cytochrome c aggregates into fibrillar structures. Characterization by transmission electron mi- croscopy and thioflavin-T binding assays shows that these species posses the characteristics of fibrils associated with the family of amyloid diseases. Our findings indicate that destabilization of the native fold of this highly α-helical protein can lead to its polymerization into β-sheet rich structures and suggest that this process does not depend on the population of partially folded monomeric states with extensive β-sheet structure. Keywords: Amyloid formation, cytochrome c, protein misfolding, protein denaturation, helical proteins Abbreviations: CD = circular dichroism; FTIR = Fourier-transform infrared. 1. Introduction The deposition of amyloid fibrils has been linked to a variety of slow-onset degenerative diseases, such as Alzheimer’s disease, senile systemic amyloidosis, Parkinson’s disease, dyalisis-related amyloi- dosis, and transmissible spongiform encephalopathies [1–4]. The proteins responsible for these diseases do not share structural or sequential identities [5]. In spite of this diversity, all amyloid fibrils display similar structural features, exhibiting a cross-β structure. In the last few years, proteins unrelated to any known human disease have been found to co