National Institute for Research and Development of Isotopic and Molecular Technologies, 67 - 103 Donat, 400293 Cluj-Napoca, Romania.
National Institute for Research and Development of Isotopic and Molecular Technologies, 67 - 103 Donat, 400293 Cluj-Napoca, Romania.
Spectrochim Acta A Mol Biomol Spectrosc. 2018 Feb 15;191:226-232. doi: 10.1016/j.saa.2017.10.032. Epub 2017 Oct 10.
A quantitative analysis of the interaction between zidovudine (AZT) and human serum albumin (HSA) was achieved using Isothermal titration calorimetry (ITC) in combination with fluorescence and H NMR spectroscopy. ITC directly measure the heat during a biomolecular binding event and gave us thermodynamic parameters and the characteristic association constant. By fluorescence quenching, the binding parameters of AZT-HSA interaction was determined and location to binding site I of HSA was confirmed. Via T NMR selective relaxation time measurements the drug-protein binding extent was evaluated as dissociation constants K and the involvement of azido moiety of zidovudine in molecular complex formation was put in evidence. All three methods indicated a very weak binding interaction. The association constant determined by ITC (3.58×10M) is supported by fluorescence quenching data (2.74×10M). The thermodynamic signature indicates that at least hydrophobic and electrostatic type interactions played a main role in the binding process.
采用等温滴定量热法（ITC）结合荧光和 H 核磁共振波谱法对叠氮胸苷（AZT）与人血清白蛋白（HSA）的相互作用进行了定量分析。ITC 直接测量生物分子结合事件过程中的热量，并提供了热力学参数和特征结合常数。通过荧光猝灭，确定了 AZT-HSA 相互作用的结合参数，并证实了其结合位点 I。通过 T1 NMR 选择性弛豫时间测量，评估了药物-蛋白质结合程度，得到解离常数 K，并证明了 AZT 中叠氮基团在分子复合物形成中的参与。这三种方法都表明存在非常弱的结合相互作用。ITC 测定的结合常数（3.58×10^4 M^-1）得到了荧光猝灭数据（2.74×10^4 M^-1）的支持。热力学特征表明，至少疏水相互作用和静电相互作用在结合过程中起主要作用。
