Centre for Interdisciplinary Research in Basic Sciences, Jamia Millia Islamia, Jamia Nagar, New Delhi 110025, India.
Department of Chemistry, Biochemistry and Forensic Science, Amity School of Applied Sciences, Amity University, Haryana 122413, India.
Biomolecules. 2021 Mar 11;11(3):414. doi: 10.3390/biom11030414.
Protein aggregation and misfolding are some of the most challenging obstacles, customarily studied for their association with amyloid pathologies. The mechanism of amyloid fibrillation development is a dynamic phenomenon involving various factors such as the intrinsic properties of protein and the physical and chemical environmental conditions. The purpose of this study was to see the thermal aggregation profile of alpha-lactalbumin (α-LA) and to delineate the effect of trehalose on its aggregation profile. α-LA was subjected to thermal aggregation at high concentrations. UV-Vis spectroscopy, a turbidity assay, intrinsic fluorescence, Rayleigh scattering and a thioflavin T (ThT) assay explained the steady outcomes that 1 M trehalose repressed α-LA aggregation in the most effective way followed by 0.75 M and 0.5 M and to a significantly lesser degree by 0.25 M. Multi spectroscopic obser Sania ations were further entrenched by microscopy. Transmission electron microscopy confirmed that in the presence of its higher concentration, trehalose hinders fibril development in α-LA. In vitro studies were further validated by in silico studies. Molecular docking analysis indicated that trehalose occupied the binding pocket cavity of α-LA and offered several significant interactions, including H-bonds with important residues. This study provides a platform for trehalose in the therapeutic management of protein aggregation-related diseases.
蛋白质聚集和错误折叠是最具挑战性的障碍之一，通常研究它们与淀粉样变病理学的关系。淀粉样纤维发展的机制是一个动态现象，涉及蛋白质的固有性质以及物理和化学环境条件等各种因素。本研究的目的是观察α-乳白蛋白（α-LA）的热聚集特性，并阐明海藻糖对其聚集特性的影响。α-LA 在高浓度下进行热聚集。紫外-可见光谱、浊度测定、内源荧光、瑞利散射和硫黄素 T（ThT）测定解释了稳定的结果，即 1 M 海藻糖以最有效的方式抑制α-LA 聚集，其次是 0.75 M 和 0.5 M，0.25 M 的抑制作用要小得多。多光谱观察进一步通过显微镜得到证实。透射电子显微镜证实，在较高浓度的海藻糖存在下，海藻糖阻碍了α-LA 中纤维的发展。体外研究进一步通过计算机模拟研究得到验证。分子对接分析表明，海藻糖占据了α-LA 的结合口袋腔，并提供了几个重要的相互作用，包括与重要残基的氢键。本研究为海藻糖在治疗与蛋白质聚集相关的疾病方面提供了一个平台。
