光谱法研究灿烂甲酚蓝与血浆蛋白质结合性质及蛋白质构象变化  

Spectroscopic Studies on the Binding Characteristics of Brilliant Cresyl Blue to Plasma Protein and the Changes of Protein Conformation

作　　者：王新[1,2,3] 周玲玲 郝俊旭 王晓芳 徐亮 刘彬[1,2,3] WANG Xin;ZHOU Ling-ling;HAO Jun-xu;WANG Xiao-fang;XU Liang;LIU Bin(School of Pharmaceutical Sciences,Liaoning University;Liaoning Provincial Engineering Research Center for Natural Products Pharmaceutica;Liaoning Provincial Key Laboratory of New Drug R＆;Shenyang 110036,China)

机构地区：[1]辽宁大学药学院,辽宁沈阳110036 [2]辽宁省天然产物制药工程技术研究中心,辽宁沈阳110036 [3]辽宁省新药研发重点实验室,辽宁沈阳110036

出　　处：《辽宁大学学报（自然科学版）》2018年第2期144-153,共10页Journal of Liaoning University：Natural Sciences Edition

基　　金：国家自然科学基金(21402125);辽宁大学"大学生创新创业训练计划"项目(x201710140291)

摘　　要：在模拟的生理条件下,采用荧光发射光谱、紫外-可见分光光度法、同步荧光光谱、三维荧光光谱和圆二色(Circular dichroism,CD)光谱等技术研究了灿烂甲酚蓝(Brilliant cresyl blue,BCB)与人血清白蛋白(Human serum albumin,HSA)的结合性质以及药物的结合对HSA构象的影响.结果表明,BCB和能猝灭HSA的内源荧光,BCB对HSA的荧光猝灭是形成HSA-BCB复合物的静态猝灭过程,紫外-可见吸收光谱的结果也进一步证实了BCB对HSA荧光的静态猝灭.BCB与HSA分子Trp残基的距离r小于7 nm,说明HSA与BCB之间能够发生非辐射能量转移,但由于r>R0值,这说明形成HSA-BCB复合物导致的静态猝灭是HSA荧光猝灭的主要原因.热力学参数计算结果表明BCB与HSA相互作用形成HSA-BCB复合物的过程是自发进行的,疏水作用力和氢键在形成HSA-BCB复合物的过程中发挥主要作用.同步荧光光谱、三维荧光光谱和CD光谱的结果说明BCB与HSA的相互作用形成的HSA-BCB复合物导致了HSA的构象发生了变化.Herein,the binding characteristics of brilliant cresyl blue（ BCB） tohuman serum albumin（ HSA） and the influences of its binding to HSA on the conformation of HSA were studiedby means of fluorescence,ultraviolet-visible（ UV-vis） absorption,synchronous fluorescence,three dimensional（ 3 D） fluorescence and circular dichroism（ CD） spectra under simulated physiological conditions. The results are as follows： The intrinsic fluorescence of HSA was quenched remarkably by BCB. The fluorescence quenching were static quenching and initiated by the formation of HSABCB complex,which were further confirmed by UV-vis absorption spectra measurements. The value of the binding distance（ r） of BCB and the tryptophan（ Trp） residues of HSA is less than 7 nm,which suggest that the energy transfer from HSA to BCB may occur. Because the value of r is greater than that of R0,the result indicates that the static quenching due to the formation of HSABCB complex is the main reason for the fluorescence quenching of HSA. Thecalculated results of thermodynamic parameters indicate that the formation processes of HSA-BCB complex isspontaneous and both hydrophobic interactions and hydrogen bonding play major roles in the formationof HSABCBcomplex. The results of synchronous fluorescence,3 D fluorescence and CD spectra suggest that the formationof HSA-BCBcomplex can give rise to the changes of HSA conformation.

关 键 词：光谱法 灿烂甲酚蓝 血浆蛋白质 结合性质 构象变化 

分 类 号：R965[医药卫生—药理学]