Institute of Chemistry and Biology of Membranes and Nanoobjects, Institut Européen de Chimie et Biologie (CNRS UMR 5248, Université de Bordeaux), 33600 Pessac, France.
Centre de RMN à Très Hauts Champs, Institut des Sciences Analytiques (CNRS, ENS Lyon, UCB Lyon 1), 69100 Villeurbanne, France.
Methods. 2018 Apr 1;138-139:26-38. doi: 10.1016/j.ymeth.2018.03.014. Epub 2018 Apr 6.
The amyloid fold is structurally characterized by a typical cross-β architecture, which is under debate to represent an energy-favourable folding state that many globular or natively unfolded proteins can adopt. Being initially solely associated with amyloid fibrils observed in the propagation of several neurodegenerative disorders, the discovery of non-pathological (or "functional") amyloids in many native biological processes has recently further intensified the general interest invested in those cross-β supramolecular assemblies. The insoluble and non-crystalline nature of amyloid fibrils and their usually inhomogeneous appearance on the mesoscopic level pose a challenge to biophysical techniques aiming at an atomic-level structural characterization. Solid-state NMR spectroscopy (SSNMR) has granted breakthroughs in structural investigations on amyloid fibrils ranging from the assessment of the impact of polymorphism in disease development to the 3D atomic structure determination of amyloid fibrils. First landmark studies towards the characterization of atomic structures and interactions involving functional amyloids have provided new impulses in the understanding of the role of the amyloid fold in native biological functions. Over the last decade many strategies have been developed in protein isotope labelling, NMR resonance assignment, distance restraint determination and 3D structure calculation of amyloid fibrils based on SSNMR approaches. We will here discuss the emerging concepts and state-of-the-art methods related to the assessment of amyloid structures and interactions involving amyloid entities by SSNMR.
淀粉样蛋白折叠在结构上的特点是典型的交叉-β 结构，这种结构被认为是一种能量有利的折叠状态，许多球状或天然无规的蛋白质都可以采用这种状态。淀粉样蛋白最初仅与在几种神经退行性疾病的传播过程中观察到的淀粉样纤维有关，但最近在许多天然生物过程中发现了非病理性（或“功能性”）淀粉样蛋白，这进一步加剧了人们对这些交叉-β 超分子组装体的普遍兴趣。淀粉样纤维的不溶性和非晶态性质以及它们在介观水平上通常不均匀的外观，对旨在进行原子水平结构表征的生物物理技术提出了挑战。固态 NMR 光谱学（SSNMR）在淀粉样纤维的结构研究方面取得了突破，从评估多态性对疾病发展的影响到淀粉样纤维的 3D 原子结构测定。首次对涉及功能性淀粉样蛋白的原子结构和相互作用进行特征描述的标志性研究，为理解淀粉样蛋白折叠在天然生物功能中的作用提供了新的动力。在过去的十年中，已经开发了许多基于 SSNMR 方法的蛋白质同位素标记、NMR 共振分配、距离约束确定和 3D 结构计算的策略，用于淀粉样纤维。我们将在这里讨论与通过 SSNMR 评估涉及淀粉样蛋白实体的淀粉样蛋白结构和相互作用相关的新兴概念和最新方法。
