摘要
在pH 7.40的Tris-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303K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。
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 .
出处
《光谱学与光谱分析》
SCIE
EI
CAS
CSCD
北大核心
2014年第3期762-766,共5页
Spectroscopy and Spectral Analysis
基金
国家自然科学基金项目(20675024)
河北省重点基础研究项目(10967126D)资助
关键词
牛血清白蛋白
格列吡嗪
荧光光谱
弹性散射荧光
Bovine serum albumin
Glipizide
Fluorescence spectra
Elastic scattering fluorescence spectrometry
