药效活性BVOCs物质组与β-牛乳球蛋白选择性结合的分子机制

Molecular Mechanism of Selective Binding of Pharmacodynamic Active BVOCs Substance Group with Bovineβ-Lactoglobulin

顶空固相微萃取气相色谱-质谱联用技术(headspace-solid phase microextraction/gas chromatography-mass spectrometry,HS-SPME/GC-MS)用于分析植物柚皮活性挥发物(botany volatile organic compounds,BVOCs)物质组与β-牛乳球蛋白(bovineβ-lactoglobulin,β-LG)相互作用,筛选出药效活性BVOCs物质组。通过定量回收BVOCs的方法,分析β-LG与药效活性BVOCs物质组的选择结合作用,计算结合参数。进而结合分子对接及光谱法建立BVOCs与β-LG结合的分子模型,探讨组学角度下药效活性BVOCs物质组与β-LG的分子作用机制。结果表明,HS-SPME/GC-MS技术能够通过β-LG与BVOCs物质组的结合作用,筛选出药效活性BVOCs物质组D-柠檬烯(dipentene,Dt)、乙酸芳樟酯(linalylacetate,La)及圆柚酮(nootkatone,Nt)。参数计算表明,β-LG与Nt的亲和力最强,但结合力不强,对La的亲和力最弱。β-LG对Dt的亲和力较弱,但结合力最强,结合率达54.66%,说明β-LG与药效活性BVOCs物质组的选择结合强度取决于BVOCs分子的化学结构特性,β-LG更倾向结合含有羰基氧结构的醛酮类BVOCs分子。本文还建立了药效活性BVOCs物质组与β-LG的分子模型,评估了BVOCs物质组(Dt,La,Nt)在β-LG分子上的结合位置。并阐明了药效活性BVOCs物质组导入后引起的β-LG二级结构的松动、伸展及构象改变是范德华力、疏水作用和氢键共同作用的结果。本研究为从BVOCs物质组角度筛选药效活性BVOCs物质组提供新方法,并为从组学角度考察药效活性BVOCs物质组与功能蛋白质分子的结合机制提供有益参考。

Headspace-solid phase microextraction/gas chromatography-mass spectrometry(HS-SPME/GC-MS)were used to analyze the interaction between theβ-lactoglobulin(β-LG)and the botany volatile organic compounds(BVOCs)from pomelo peel to screen out the pharmacodynamic active BVOCs substance group.The selective binding effect betweenβ-LG and BVOCs was analyzed by quantitative recovery of BVOCs,and the binding parameters were calculated.Then,the molecular model of BVOCs binding withβ-LG was established by molecular docking and spectroscopic method,and the molecular mechanism of interaction between pharmacodynamic active BVOCs andβ-LG was discussed from the perspective of omics.The results showed that dipentene(Dt),linalylacetate(La)and nootkatone(Nt)of BVOCs were selected by HS-SPME/GC-MS by the interaction ofβ-LG and BVOCs substance group.Parameter calculation showed thatβ-LG had the strongest affinity with Nt,but the binding force was not strong,and the affinity for La was the weakest.The affinity ofβ-LG to Dt was weak,but the binding force was the strongest,with a binding rate of 54.66%,indicating that the selective binding strength ofβ-LG with the pharmacodynamic active BVOCs depended on the chemical structure of BVOCs molecules.Theβ-LG preferred to bind to the aldehyde and ketone BVOCs molecules containing carbonyl oxygen structure.The molecular model ofβ-LG and BVOCs group(Dt,La,Nt)was established to evaluate the binding position of BVOCs group(Dt,La,Nt)onβ-LG.The loosening,extension and conformational change ofβ-LG secondary structure caused by the introduction of BVOCs are the result of van der Waals force,hydrophobicity and hydrogen bonding.This study provides a new method for screening pharmacodynamic active BVOCs from the perspective of whole substance group of BVOCs,and provides a useful reference for investigating the binding mechanism between pharmacodynamic active BVOCs and functional protein molecules from the perspective of omics.