spectroscopic analysis of amyloid fibril formation in sh3？domains淀粉样原纤维形成的光谱分析在sh3 域.pdfVIP

Spectroscopy 17 (2003) 647–652 647 IOS Press Spectroscopic analysis of amyloid fibril formation in SH3-domains Salvador Ventura ∗ and Luis Serrano European Molecular Biology Laboratory, Meyerhofstrasse 1, D-69117 Heidelberg, Germany Abstract. The aggregation of proteins in fibrillar form is a problem of critical importance in a wide range of abnormal disease states. To decipher the molecular mechanism of formation of protein fibrillar aggregates we have chosen to study as model SH3 domains that exhibit different abilities to polymerize into amyloid fibrils. While being not related to any known disease, the SH3 domain of the p85α subunit of phosphatidylinositol 3 kinase has been found to form amyloid fibrils in vitro under acidic conditions, meanwhile, we have found that the spectrin SH3-domain, sharing the same fold and some sequential identity keeps its native conformation under the same conditions. The use of spectroscopic methods to study these properties is illustrated in the present job, and correlated to direct sample observation by electron microscopy. Keywords: Amyloid formation, UV, fluorescence, circular dichroism, SH3 domain 1. Introduction Amyloidoses are a group of protein misfolding diseases that are characterised by the polymerisation of normally innocuous and soluble proteins or peptides into insoluble proteinaceous deposits. Among these diseases, we can point out the transmissible spongiform encephalopathies, Alzheimer’s disease and type II diabetes [1]. The proteins responsible for these diseases do not share structural or sequential identities [2]. In spite of this diversity, all amyloid fibrils display similar features that can be checked by different spectroscopic methods: (1) they are long, straight and unbranched fibrils with a diameter between 40–120 Å; (2) they bind to dyes such Congo Red and thioflavin-T, and (3) X-r