Recombinant amyloidogenic domain of ApoA-I: Analysis of its fibrillogenic potential
- 作者：Sonia Di Gaetano ; Fulvio Guglielmi ; Angela Arciello ; Palma Mangione ; Maria Monti ; Daniela Pagnozzi ; Sara Raimondi ; Sofia Giorgetti ; Stefania Orrù ; Claudio Canale et al.
- 刊名：Biochemical and Biophysical Research Communications
- 出版年：2006
- 期刊代码：159_0006291x
- 类别：bio
- 出版时间：8 December 2006
- 卷：351
- 期：1
- 页码：223-228
- 文件大小：590 K
摘要
A variety of amyloid diseases are associated with fibrillar aggregates from N-terminal fragments of ApoA-I generated through a largely unexplored multi-step process. The understanding of the molecular mechanism is impaired by the lack of suitable amounts of the fibrillogenic polypeptides that could not be produced by recombinant methods so far. We report the production and the conformational analysis of recombinant ApoA-I 1–93 fragment. Similarly to the polypeptide isolated ex vivo, a pH switch from 7 to 4 induces a fast and reversible conformational transition to a helical state and leads to the identification of a key intermediate in the fibrillogenesis process. Limited proteolysis experiments suggested that the C-terminal region is involved in helix formation. The recombinant polypeptide generates fibrils at pH 4 on a time scale comparable with that of the native fragment. These findings open the way to studies on structural, thermodynamic, and kinetic aspects of ApoA-I fibrillogenesis.