Amyloids are proteinaceous deposits considered an underlying pathological hallmark of several degenerative diseases. The mechanism of amyloid formation and its inhibition still represent challenging issues, especially when protein structure cannot be investigated by classical biophysical techniques as for the intrinsically disordered proteins (IDPs). In this view, the need to find an alternative way for providing molecular and structural information regarding IDPs prompted us to set a novel, to our knowledge, approach focused on UV Resonance Raman (UVRR) spectroscopy. To test its applicability, we study the fibrillation of hen-egg white lysozyme (HEWL) and insulin as well as their interaction with resveratrol, employing also intrinsic fluorescence spectroscopy, Fourier transform infrared (FTIR) spectroscopy, and atomic force microscopy (AFM). The increasing of the β-sheet structure content at the end of protein fibrillation probed by FTIR occurs simultaneously with a major solvent exposure of tryptophan (Trp) and tyrosine (Tyr) residues of HEWL and insulin, respectively, as revealed by UVRR and intrinsic fluorescence spectroscopy. However, because the latter technique is successfully used when proteins naturally contain Trp residues, it shows poor performances in the case of insulin, and the information regarding its tertiary structure is exclusively provided by UVRR spectroscopy. The presence of an increased concentration of resveratrol induces mild changes in the secondary structure of both protein fibrils while remodeling HEWL fibril length and promoting the formation of amorphous aggregates in the case of insulin. Although the intrinsic fluorescence spectra of proteins are hidden by resveratrol signal, UVRR Trp and Tyr bands are resonantly enhanced, showing a good sensitivity to the presence of resveratrol and marking a modification in the noncovalent interactions in which they are involved. Our findings demonstrate that UVRR is successfully employed in the study of aggregation-prone proteins and of their interaction with ligands, especially in the case of Trp-lacking proteins.

淀粉样蛋白是蛋白质沉积物，被认为是几种退行性疾病的潜在病理标志。淀粉样蛋白形成的机制及其抑制仍然是具有挑战性的问题，特别是当蛋白质结构无法通过经典生物物理技术研究时，如内在无序蛋白质 (IDP)。在这种观点中，需要找到一种替代方法来提供有关 IDP 的分子和结构信息，这促使我们设置了一种新的方法，据我们所知，该方法专注于紫外共振拉曼 (UVRR) 光谱。为了测试其适用性，我们研究了鸡蛋清溶菌酶 (HEWL) 和胰岛素的纤颤以及它们与白藜芦醇的相互作用，还采用了本征荧光光谱、傅里叶变换红外 (FTIR) 光谱和原子力显微镜 (AFM)。β如 UVRR 和固有荧光光谱所揭示的，FTIR 探测到的蛋白质原纤维化末端的片层结构含量与 HEWL 和胰岛素的色氨酸 (Trp) 和酪氨酸 (Tyr) 残基的主要溶剂暴露同时发生。然而，由于后一种技术在蛋白质天然含有色氨酸残基时成功使用，它在胰岛素的情况下表现不佳，并且有关其三级结构的信息仅由 UVRR 光谱提供。白藜芦醇浓度增加会导致两种蛋白质原纤维的二级结构发生轻微变化，同时重塑 HEWL 原纤维长度并在胰岛素的情况下促进无定形聚集体的形成。虽然蛋白质的内在荧光光谱被白藜芦醇信号隐藏，UVRR Trp 和 Tyr 带共振增强，对白藜芦醇的存在表现出良好的敏感性，并标志着它们所涉及的非共价相互作用发生了改变。我们的研究结果表明，UVRR 成功地用于研究易于聚集的蛋白质及其与配体的相互作用，特别是在缺乏色氨酸的蛋白质的情况下。