The study on the interaction between human serum albumin and a new reagent with antitumour activity by spectrophotometric methods
The study on the interaction between human serum albumin and a new reagent with antitumour activity by spectrophotometric methods
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DOI：
10.1016/j.jphotochem.2004.05.017
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发表时间：
2004-10-01
影响因子：
4.3
通讯作者：
Hu, ZD
中科院分区：
文献类型：
--
作者：
Gao, H;Lei, LD;Hu, ZD
本文首次在模拟生理条件下，采用荧光光谱结合紫外-可见吸收光谱、圆二色谱（CD）和傅立叶变换红外（FT-IR）光谱技术研究了抗肿瘤新试剂2-羟基-3-硝基-9-芴酮（HNF）与人血清白蛋白（HSA）的结合。HNF对HSA有较强的荧光猝灭作用，根据Stern-Volmer（S-V）方程推测其猝灭机理为静态猝灭。计算出300、310和320 K下HNF与HSA的结合常数分别为6.08 x 10（5）、3.80 x 10（5）和2.79 x 10（5）M-1，相应的结合位点数（n）分别为1.1、1.0和1.0。实验结果表明，HNF与HSA的结合引起了HSA构象的变化。CD谱的定量分析数据从游离HSA中α-螺旋的60.3%到HNF-HSA复合物中的56.5%进一步证实了HNF修饰了蛋白质的二级结构。热力学参数Δ H度和Δ S度分别为-31.10 kJ mol（-1）和6.87 J mol（-1）K-1，表明疏水力在HNF与HSA的相互作用中起主要作用.这些实验结果和理论数据表明，HNF能够与HSA结合，并在体内有效转运和消除，为进一步的药物设计提供了指导。(C)2004 Elsevier B. V.保留所有权利。
In this work, the binding of 2-hydroxy-3-nitro-9-fluorenone (HNF; a new reagent with antitumour activity) to human serum albumin (HSA) was investigated by fluorescence spectroscopy combined with UV-Vis absorption, circular dichroism (CD), and Fourier transform infrared (FT-IR) spectrophotometric techniques under simulative physiological conditions for the first time. A strong fluorescence quenching reaction of HNF to HSA was observed and the quenching mechanism was suggested as static quenching according to the Stern-Volmer (S-V) equation. The binding constants of HNF with HSA at 300, 3 10 and 320 K were calculated as 6.08 x 10(5), 3.80 x 10(5) and 2.79 x 10(5) M-1, respectively, and corresponding numbers of binding sites (n) were 1.1, 1.0 and 1.0. Experimental results observed showed that the binding of HNF to HSA induced conformational change of HSA. The quantitative analysis data of CD spectra from that of the alpha-helix 60.3% in free HSA to 56.5% in the HNF-HSA complex further confirmed that the secondary structure of the protein was modified by HNF. The thermodynamic parameters, standard enthalpy change (DeltaHdegrees) and the standard entropy change (DeltaSdegrees), were obtained to be -31.10 kJ mol(-1) and 6.87 J mol(-1) K-1, respectively, which indicated that a hydrophobic force played a major role in the interaction of HNF with HSA. All these experimental results and theoretical data clarified that HNF could bind to HSA and be effectively transported and eliminated in body, which could be a useful guideline for further drug design. (C) 2004 Elsevier B.V. All rights reserved.