Interaction of virstatin with human serum albumin: spectroscopic analysis and molecular modeling.
Interaction of virstatin with human serum albumin: spectroscopic analysis and molecular modeling.
复制标题
DOI：
10.1371/journal.pone.0037468
复制
发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Chakrabarti P
中科院分区：
文献类型：
--
作者：
Chatterjee T;Pal A;Dey S;Chatterjee BK;Chakrabarti P
Virstatin是一种抑制霍乱弧菌毒力调节的小分子，霍乱弧菌是霍乱的病原体。在这里，我们用各种生物物理方法报道了韦他汀与人血清白蛋白（HSA）的相互作用。利用不同异构体形式的HSA （N型~ pH 7.2， B型~ pH 9.0和F型~ pH 3.5）通过吸收和荧光光谱监测药物结合。在与药物结合时，HSA的固有荧光明显猝灭。福斯特型荧光共振能量转移（FRET）得到的供体（HSA中的Trp214）与受体（virstatin）之间的距离r为3.05 nm。ITC数据表明，N和B异构体的结合是一个焓驱动的过程，其结合常数K a分别为6.09×105 M−1和4.47×105 M−1。利用圆二色性（CD）和傅里叶变换红外光谱（FTIR）研究了HSA与药物相互作用后的构象变化。当蛋白质与药物的摩尔比为1∶1时，远紫外CD光谱显示，所有构象的α-螺旋度都增加，蛋白质对尿素和热展开具有稳定性。分子对接研究揭示了可能参与蛋白-药物相互作用的残基，并表明virstatin与Site I（亚结构域IIA）结合，也称为华法林结合位点。
Virstatin is a small molecule that inhibits Vibrio cholerae virulence regulation, the causative agent for cholera. Here we report the interaction of virstatin with human serum albumin (HSA) using various biophysical methods. The drug binding was monitored using different isomeric forms of HSA (N form ∼pH 7.2, B form ∼pH 9.0 and F form ∼pH 3.5) by absorption and fluorescence spectroscopy. There is a considerable quenching of the intrinsic fluorescence of HSA on binding the drug. The distance (r) between donor (Trp214 in HSA) and acceptor (virstatin), obtained from Forster-type fluorescence resonance energy transfer (FRET), was found to be 3.05 nm. The ITC data revealed that the binding was an enthalpy-driven process and the binding constants K a for N and B isomers were found to be 6.09×105 M−1 and 4.47×105 M−1, respectively. The conformational changes of HSA due to the interaction with the drug were investigated from circular dichroism (CD) and Fourier Transform Infrared (FTIR) spectroscopy. For 1∶1 molar ratio of the protein and the drug the far-UV CD spectra showed an increase in α- helicity for all the conformers of HSA, and the protein is stabilized against urea and thermal unfolding. Molecular docking studies revealed possible residues involved in the protein-drug interaction and indicated that virstatin binds to Site I (subdomain IIA), also known as the warfarin binding site.
登录
查看更多内容
影响因子：
14.8
作者：
Greenfield, Norma J.
通讯作者：
Greenfield, Norma J.
影响因子：
7.7
作者：
Connelly, Patrick R.
通讯作者：
Connelly, Patrick R.
影响因子：
4.1
作者：
KHAN, MY
通讯作者：
KHAN, MY
影响因子：
56.9
作者：
Hung, DT;Shakhnovich, EA;Mekalanos, JJ
通讯作者：
Mekalanos, JJ
影响因子：
5.6
作者：
Jones, G;Willett, P;Taylor, R
通讯作者：
Taylor, R