Instituto de Biología Molecular y Celular de Rosario, Consejo Nacional de Investigaciones Científicas y Técnicas, Universidad Nacional de Rosario, Suipacha 531, S2002LRK Rosario, Argentina.
J Inorg Biochem. 2012 Dec;117:334-41. doi: 10.1016/j.jinorgbio.2012.06.011. Epub 2012 Jun 29.
α-Synuclein (AS) aggregation is associated to neurodegeneration in Parkinson's disease (PD). At the same time, alterations in metal ion homeostasis may play a pivotal role in the progression of AS amyloid assembly and the onset of PD. Elucidation of the structural basis directing AS-metal interactions and their effect on AS aggregation constitutes a key step towards understanding the role of metal ions in AS amyloid formation and neurodegeneration. Despite of the reported evidences that link Zn(2+) with the pathophysiology of PD and the fact that this metal ion was shown to promote AS fibrillation in vitro, neither the structural characterization of the binding sites nor the identification of the amino acids involved in the interaction of Zn(2+) with the protein AS has been carried out. By using NMR spectroscopy, we have addressed here unknown structural details related to the binding of Zn(2+) to the protein AS through the design of site-directed and domain truncated mutants of AS. The binding of zinc to the Aβ peptide was also studied and discussed comparatively. Although the results of this study contribute to the understanding of the structural and molecular basis behind the acceleration of AS fibrillation mediated by Zn(2+), the low affinity that characterizes the interaction of Zn(2+) with AS contrasts strongly with the high-affinity features reported for the binding of this metal ion to other target proteins linked to human amylodosis such as Aβ peptide and the Islet Amyloid Polypeptide (IAPP), challenging the biological relevance of zinc interactions in the pathogenesis of PD.
α-突触核蛋白(AS)聚集与帕金森病(PD)的神经退行性变有关。与此同时,金属离子动态平衡的改变可能在 AS 淀粉样蛋白组装的进展和 PD 的发病中起着关键作用。阐明指导 AS-金属相互作用的结构基础及其对 AS 聚集的影响,是理解金属离子在 AS 淀粉样形成和神经退行性变中的作用的关键步骤。尽管有报道称锌(Zn(2+)与 PD 的病理生理学有关,并且该金属离子已被证明在体外促进 AS 纤维形成,但尚未对结合位点的结构特征进行表征,也未鉴定与 Zn(2+)与 AS 蛋白相互作用相关的氨基酸。通过使用 NMR 光谱,我们通过设计 AS 的定点和结构域截断突变体,解决了与 Zn(2+)与蛋白质 AS 结合相关的未知结构细节。还比较研究了锌与 Aβ肽的结合。尽管这项研究的结果有助于理解 Zn(2+)介导的 AS 纤维加速的结构和分子基础,但 Zn(2+)与 AS 的相互作用的低亲和力与该金属离子与其他与人类淀粉样变性相关的靶蛋白(如 Aβ肽和胰岛淀粉样多肽(IAPP))的高亲和力特征形成鲜明对比,挑战了锌相互作用在 PD 发病机制中的生物学相关性。