CAJ | 학술논문

目的：研究炎琥宁（YHN）与牛血清白蛋白（BSA）之间的相互作用，为后续琥宁类药物的研发和进一步探讨炎琥宁在生物体内与蛋白质的作用机制提供理论依据。方法：在优化的实验条件下，运用荧光猝灭光谱法、紫外光谱法和同步荧光光谱研究炎琥宁与BSA的相互作用。283.15 K、298.15 K和313.15 K温度下，根据 S-V方程计算出猝灭常数（KSV）和速率常数（Kq）；根据L-B双倒数方程计算出静态猝灭结合常数（KLB）；根据双对数方程计算出结合常数（Kb）和结合位点数（n）；根据热力学公式计算出焓变（ΔH）、熵变（ΔS）和吉布斯自由能变（ΔG）；根据Hill方程计算出 Hill系数（nH）。结果：3个温度下的BSA荧光强度随着炎琥宁浓度升高，有规律地降低；KSV、Kq、KLB、Kb、n和nH值随着温度升高而降低；ΔG<0，ΔH<0，ΔS<0；n约等于1；nH大于1。结论：炎琥宁与BSA形成基态复合物从而猝灭BSA的内源性荧光，猝灭机理为静态猝灭。通过计算反应热力学参数值，可推断炎琥宁与BSA作用力主要为氢键和范德华力。炎琥宁与BSA可形成一个结合位点，表明炎琥宁与BSA之间具有一定的结合作用，炎琥宁在体内可被蛋白质储存和转运。生成自由能变ΔG为负值，表明炎琥宁与BSA的作用过程是一个自发过程。nH大于1，表明炎琥宁有正协同作用。两者的结合部位主要位于亚螺旋域ⅡA中。同步荧光光谱表明炎琥宁对BSA构象产生一定的影响，使BSA腔内疏水环境的极性减弱，结合位点更接近于酪氨酸。
목적：연구염호저（YHN）여우혈청백단백（BSA）지간적상호작용，위후속호저류약물적연발화진일보탐토염호저재생물체내여단백질적작용궤제제공이론의거。방법：재우화적실험조건하，운용형광졸멸광보법、자외광보법화동보형광광보연구염호저여BSA적상호작용。283.15 K、298.15 K화313.15 K온도하，근거 S-V방정계산출졸멸상수（KSV）화속솔상수（Kq）；근거L-B쌍도수방정계산출정태졸멸결합상수（KLB）；근거쌍대수방정계산출결합상수（Kb）화결합위점수（n）；근거열역학공식계산출함변（ΔH）、적변（ΔS）화길포사자유능변（ΔG）；근거Hill방정계산출 Hill계수（nH）。결과：3개온도하적BSA형광강도수착염호저농도승고，유규률지강저；KSV、Kq、KLB、Kb、n화nH치수착온도승고이강저；ΔG<0，ΔH<0，ΔS<0；n약등우1；nH대우1。결론：염호저여BSA형성기태복합물종이졸멸BSA적내원성형광，졸멸궤리위정태졸멸。통과계산반응열역학삼수치，가추단염호저여BSA작용력주요위경건화범덕화력。염호저여BSA가형성일개결합위점，표명염호저여BSA지간구유일정적결합작용，염호저재체내가피단백질저존화전운。생성자유능변ΔG위부치，표명염호저여BSA적작용과정시일개자발과정。nH대우1，표명염호저유정협동작용。량자적결합부위주요위우아라선역ⅡA중。동보형광광보표명염호저대BSA구상산생일정적영향，사BSA강내소수배경적겁성감약，결합위점경접근우락안산。
The interaction betweenYan-Hu-Ning (YHN) and bovine serum albumin (BSA) was investigated in order to provide further theoretical evidences on action mechanism study between YHN and proteins within the organism. Under optimal conditions, the interaction between YHN and BSA was studied by fluorescence quenching, ultraviolet absorption spectrometry and synchronous fluorescence spectroscopy. At the temperature of 283.15 K, 298.15 K and 313.15 K, quenching constant (KSV) and speed constant (Kq) were calculated by S-V curves. Static quenching constant (KLB) was obtained by L-B double reciprocal equation. Double logarithmic equation was used to calculate the binding constants (Kb) and the number of binding site (n). Thermodynamic equation was used to obtainΔH,ΔS,ΔG. Hill’s coefficients (nH) was obtained by Hill equation. The results showed that at three different temperatures, along with the increasing of YHN concentration, the fluorescence intensity of BSA decreased regularly. The value of KSV, Kq, KLB, Kb, n and nH decreased with the increasing of temperature;ΔG < 0,ΔH < 0,ΔS < 0; n was approximately equal to 1; nH > 1. It was concluded that YHN-BSA complex quenched the intrinsic fluorescence of BSA. The mechanism of fluorescence quenching was static quenching. The main binding forces were deduced as hydrogen bonds and Van der Waals forces from calculated values of thermodynamic parameters. YHN and BSA can form a binding site, which indicated certain binding interaction between YHN and BSA. YHN can be stored and transported by protein within the body. Free energy was produced to transformΔG into negative value. It showed that the process of binding between YHN and BSA was spontaneous. The nH was more than 1. It indicated that YHN had positive cooperative effect. The primary binding site was located at subdomainⅡA. The synchronous fluorescence spectra showed certain influence on the conformation of BSA by YHN. It led to the weakening of polarity within BSA and the binding site to be closer to the tyrosine.