A novel method for the study of molecular interaction by using microscale thermophoresis
A novel method for the study of molecular interaction by using microscale thermophoresis
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DOI：
10.1016/j.talanta.2014.09.038
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发表时间：
2015-01-15
期刊：
影响因子：
6.1
通讯作者：
Harrington, Perter de B.
中科院分区：
文献类型：
--
作者：
Mao, Yexuan;Yu, Lanlan;Harrington, Perter de B.
蛋白质及其伴侣结合事件的基础研究是药物开发的关键。本研究采用微尺度热电泳技术研究了有机染料异硫氰酸荧光素（FITC）与牛血清白蛋白（BSA）的分子相互作用，并与常规荧光光谱分析结果进行了比较。MST数据表明，在较短的相互作用时间内，FITC通过弱相互作用而不是标记蛋白，对BSA表现出较高的结合亲和力。通过华法林和布洛芬作为BSA位点标记的竞争策略，证明FITC主要结合在BSA II位点的疏水口袋上，而不是BSA I位点。除了结合亲和力外，MST还提供了关于BSA聚集和FITC与BSA聚集体结合的额外信息，这是荧光光谱无法获得的。这项工作证明了MST作为一种新的研究蛋白质-小分子相互作用的方法是强大和可靠的。(C) 2014 Elsevier B.V.版权所有
The fundamental studies for the binding events of protein and its partner are crucial in drug development. In this study, a novel technology named microscale thermophoresis (MST) was applied in the investigation of molecular interaction between an organic dye fluorescein isothiocyanate (FITC) and bovine serum albumin (BSA), and the results were compared with those obtained from conventional fluorescence spectroscopy. The MST data demonstrated that with a short interaction time, FITC showed a high binding affinity for BSA by weak interaction instead of labeling the protein. By using competitive strategies in which warfarin and ibuprofen acted as the site markers of BSA, FITC was proven to mainly bind to the hydrophobic pocket of site II of BSA compared to site I of BSA. Except for the binding affinity, MST also provided additional information with respect to the aggregation of BSA and the binding of FITC to BSA aggregates, which is unobtainable by fluorescence spectroscopy. This work proves that MST as a new approach is powerful and reliable for investigation of protein-small molecule interaction. (C) 2014 Elsevier B.V. All rights reserved.