目的:研究阿霉素(doxorubicin,DOX)与八肋游仆虫中心蛋白的N-端部分(N-terminal domain of Euplotes oc-tocarinatus centrin,N-EoCen)的相互作用.方法:采用圆二色光谱以及三维荧光光谱法研究N-EoCen的构象变化,采用稳态荧光光谱法与等温滴定法(ITC)研究DOX与N-EoCen的作用机制,采用分子对接技术模拟DOX与N-EoCen的结合位点.结果:N-EoCen与DOX结合后,构象发生变化,α-helix含量减少.DOX以摩尔比1∶1与N-EoCen结合,结合常数约为105 L/mol.该反应为放热反应,熵变对自由能的贡献较小,反应主要由焓变驱动.结论:DOX主要通过疏水作用、范德华力以及氢键等多种作用与N-EoCen结合,从而使N-EoCen的构象发生改变.
Study of the interaction between doxorubicin and the centrin by spectroscopy,calorimetry and molecular docking techniques
Objective:To study the interaction between doxorubicin(DOX)and the N-terminal domain of Euplotes octocarinatus centrin(N-EoCen).Methods:Circular dichroism spectroscopy and three-dimensional fluorescence spec-troscopy were used to study the conformational change of N-EoCen.Steady-state fluorescence spectroscopy and isothermal titration calorimetry(ITC)were used to study the interaction mechanism between DOX and N-EoCen.Molecular docking technology was used to simulate the binding site between DOX and N-EoCen.Results:When N-EoCen bound to DOX,the conformation changed and theα-helix content decreased.DOX was bound to N-EoCen at a molar ratio of 1∶1 with a binding constant of about 105 L/mol.The reaction was exthermic,the contribution of entropy change to the free energy was small,and the reaction was mainly driven by enthalpy change.Conclusion:DOX binds to N-EoCen mainly through various inter-actions such as hydrophobic interaction,van der Waals force and hydrogen bonding,thereby changing the conformation of N-EoCen.
NETL
NSTL
万方数据
维普
