The proteins, which bind albuminspecific dye Evans blue, are revealed in the low-molecular protein fraction of the blood serum from Scorpaena porcus L. and identified as serum albumin. They were represented by three b...The proteins, which bind albuminspecific dye Evans blue, are revealed in the low-molecular protein fraction of the blood serum from Scorpaena porcus L. and identified as serum albumin. They were represented by three bands in 2D-SDS-PAAG. MALDI-TOF-analysis revealed the fundamental similarity of the mass spec-trum of the fragments of tryptic cleavage of proteins with molecular weight 73 and 76 kDa. The role of du-plications and intragenic reconstructions in the creation of the plurality of scorpaena albumins is discussed.展开更多
文摘The proteins, which bind albuminspecific dye Evans blue, are revealed in the low-molecular protein fraction of the blood serum from Scorpaena porcus L. and identified as serum albumin. They were represented by three bands in 2D-SDS-PAAG. MALDI-TOF-analysis revealed the fundamental similarity of the mass spec-trum of the fragments of tryptic cleavage of proteins with molecular weight 73 and 76 kDa. The role of du-plications and intragenic reconstructions in the creation of the plurality of scorpaena albumins is discussed.
基金National Natural Science Foundation of China(31360371)the Key Research and Development Project of Jiangxi Province(20181BBH80010,20192ZDF04032,20202BBF63012)The Science Foundation of the Education Department of Jiangxi Province(GJJ201821,GJJ 180918)。
文摘实验室前期研究结果表明,在国家规定的药食两用物品名单中,丁香抗氧化活性最强。前人发现天然产物的抗氧化功能与抗糖基化活性息息相关。因此,目的是在此基础上进一步对丁香精油(clove essential oil,CEO)的抗糖基化活性及其中含量最高的组分—丁香酚(Eugenol)与牛血清白蛋白(bovine serum albumin,BSA)的相互作用进行研究。在低密度脂蛋白(low density lipoprotein,LDL)的非酶糖基化孵育体系中,光谱测定结果表明CEO对LDL糖基化早期、中期和末期产物的生成均具有显著的抑制效果,且对末期产物的抑制作用最强。采用气相色谱-质谱联用技术(gas chromatography mass spectrometry,GC-MS)对CEO分析发现,CEO中含量最多的组分是丁香酚。通过多光谱和分子对接对丁香酚与BSA的相互作用研究发现,紫外-可见(ultraviolet-visible,UV-Vis)光谱表明丁香酚与BSA之间存在相互作用;在荧光发射光谱中,随着丁香酚浓度增加,BSA的荧光强度逐渐增强且发生蓝移,进一步证明了二者之间存在相互作用。通过计算不同温度下的结合常数发现,丁香酚与BSA产生相互作用过程中有热力学过程参与,热力学参数和位点标记竞争试验表明丁香酚通过氢键和范德华力与BSA在位点Ⅰ结合。随着丁香酚浓度增加,丁香酚与BSA混合体系的同步荧光(synchronous fluorescence,SF)、三维荧光(three-dimensional,3D)和傅里叶变换红外(Fourier transform infrared,FTIR)光谱的信号强度在发生变化的同时也发生了位移,表明丁香酚的添加使BSA构象发生了改变。通过分子对接技术进一步验证了丁香酚与BSA间相互作用的试验结果。该研究为丁香进一步开发应用提供理论支持。