CAJ | 학술논문

在pH 7.40的T ris-HCl缓冲溶液中，利用以蛋白荧光变化为考察对象的传统荧光光谱法和以药物荧光变化为考察对象的弹性散射荧光法，分别研究了293和303 K温度下格列吡嗪（Gli）与牛血清白蛋白（BSA）之间的反应机理，两种方法所得结论均一致。即Gli与BSA之间猝灭方式为动态猝灭；两者主要通过疏水作用力结合，结合位点主要位于BSA的疏水区，结合位点数约为1；Hill系数nH 小于1，表现为弱的负协同作用。弹性散射荧光法得到的Gli-BSA体系的结合常数均比传统荧光光谱法大，表明以药物荧光变化为考察对象的研究更准确、合理，并通过紫外光谱法对其所得结果的合理性进行了验证。结果表明：传统荧光光谱法利用蛋白为研究对象，研究药物与蛋白之间的相互作用存在一定的不足，其谱图只能反映蛋白分子与药物相互作用的部分信息，而弹性散射荧光法利用药物为研究对象，能够更全面、准确地表达药物与蛋白之间的作用信息。
재pH 7.40적T ris-HCl완충용액중，이용이단백형광변화위고찰대상적전통형광광보법화이약물형광변화위고찰대상적탄성산사형광법，분별연구료293화303 K온도하격렬필진（Gli）여우혈청백단백（BSA）지간적반응궤리，량충방법소득결론균일치。즉Gli여BSA지간졸멸방식위동태졸멸；량자주요통과소수작용력결합，결합위점주요위우BSA적소수구，결합위점수약위1；Hill계수nH 소우1，표현위약적부협동작용。탄성산사형광법득도적Gli-BSA체계적결합상수균비전통형광광보법대，표명이약물형광변화위고찰대상적연구경준학、합리，병통과자외광보법대기소득결과적합이성진행료험증。결과표명：전통형광광보법이용단백위연구대상，연구약물여단백지간적상호작용존재일정적불족，기보도지능반영단백분자여약물상호작용적부분신식，이탄성산사형광법이용약물위연구대상，능구경전면、준학지표체약물여단백지간적작용신식。
In the Tris-HCl buffer solution with pH was 7.40 ,the interaction between glipizide (Gli) and bovine serum albumin (BSA) was investigated by classical fluorescence spectroscopy with the change of protein as investigation object and elastic scat-tering fluorescence spectrometry with the change of drugs as investigation object at 293 K and 303 K ,the conclusions of the two methods were consistent .Results showed that Gli could quench the intrinsic fluorescence of BSA ,and the quenching mechanism was a dynamic quenching process .The hydrophobic force played an important role in the conjugation reaction between BSA and Gli ,the binding site mainly located in BSA hydrophobic region and the number of binding site (n) in the binary system was ap-proximately to 1 .The values of Hill’s coefficients were less than 1 ,which indicated the weak negative cooperativity in BSA-Gli system .The binding constant (Ka ) obtained by elastic scattering fluorescence spectrometric was much larger than the one ob-tained by classical fluorescence spectroscopy ,indiciating that it was more accurate and reasonable when using the change of drug ’ s fluorescence as the research object .At last ,the scientificalness of the new method based on elastic scattering fluorescence spec-trometric was verified by ultraviolet spectroscopy .The research results showed that there existed insufficiency in analysis of the interaction of drug with protein by classical fluorescence spectroscopy with the change of protein as investigation object ,and the fluorescence spectrogram only reflected partial information of the interaction between drug and protein ,while the interaction be-tween drug and protein could be better expressed by elastic scattering fluorescence spectrometry with the change of drugs as in-vestigation object .