Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, 202002, Uttar Pradesh, India.
Interdisciplinary Biotechnology Unit, Aligarh Muslim University, Aligarh, 202002, Uttar Pradesh, India.
Int J Biol Macromol. 2016 Nov;92:1220-1228. doi: 10.1016/j.ijbiomac.2016.08.035. Epub 2016 Aug 12.
In this work, binding of garlic component-Diallysulfide (DAS) with major human blood transport protein, Human Serum Albumin (HSA) and its anti- amyloidogenic behavior has been studied by utilizing various spectroscopic and molecular docking strategies. The HSA exhibit significant reduction in fluorescence intensity upon interaction with DAS. DAS quenches the fluorescence of HSA in concentration dependent manner with binding affinity of 1.14×10M. UV-visible spectroscopy results confirm the formation of DAS-HSA complex and secondary structure of HSA get stabilized upon complexation with DAS as observed by far UV CD spectroscopy and Differential Scanning Calorimetry. The topology of HSA in absence and presence of DAS was monitored through Dynamic Light Scattering (DLS) technique, inferred that protein becomes more compact in presence of DAS. Further, molecular docking study shows that DAS bind to the nearby site II in subdomain III of HSA. Moreover, effect of DAS was studied on HSA fibrillation process. ThT binding, ANS fluorescence assay, CD measurement, DLS and Transmission Electron Microscopy (TEM) results altogether confirm the anti-amyloidogenic property of DAS. This work will provide biophysical insight into the interaction of DAS with HSA and will help in designing more potential therapeutic strategies against protein aggregation by exploiting other related compounds.
在这项工作中，利用各种光谱和分子对接策略研究了大蒜成分——二烯丙基二硫（DAS）与人血清白蛋白（HSA）的结合及其抗淀粉样变性行为。HSA 与 DAS 相互作用时，其荧光强度显著降低。DAS 以浓度依赖的方式猝灭 HSA 的荧光，结合亲和力为 1.14×10M。紫外可见光谱结果证实了 DAS-HSA 复合物的形成，远紫外 CD 光谱和差示扫描量热法表明，HSA 的二级结构在与 DAS 络合后得到稳定。通过动态光散射（DLS）技术监测 HSA 在有无 DAS 存在下的拓扑结构，推断出 DAS 存在时蛋白质变得更加紧凑。此外，分子对接研究表明 DAS 结合到 HSA 亚结构域 III 中附近的 II 位点。此外，还研究了 DAS 对 HSA 纤化过程的影响。ThT 结合、ANS 荧光测定、CD 测量、DLS 和透射电子显微镜（TEM）结果共同证实了 DAS 的抗淀粉样变性特性。这项工作将为 DAS 与 HSA 的相互作用提供生物物理见解，并通过利用其他相关化合物为设计更多针对蛋白质聚集的潜在治疗策略提供帮助。
