温度、尿素、盐酸胍等因素对烟草蛋白PR-1a荧光光谱的影响研究  

The Influence of Temperature,Urea and Guanidine Hydrochloride On the Fluorescence Spectrum of Tobacco PR-1a

作　　者：文科[1] 杜林方[1] 

机构地区：[1]四川大学生命科学学院生物资源与生态环境教育部重点实验室,四川成都610064

出　　处：《四川师范大学学报（自然科学版）》2015年第5期734-739,共6页Journal of Sichuan Normal University（Natural Science）

基　　金：四川省应用技术研究与开发项目基金(2008SG0025)

摘　　要：诱导表达GST-PR-1a蛋白,利用蛋白质内源荧光和外源ANS荧光为探针,对分离的重组表达蛋白进行热稳定性、盐酸胍、尿素研究.50~80℃处理时,PR-1a内源荧光和外源荧光几乎无变化,90℃时荧光强度均有明显降低.盐酸胍、尿素处理后,随着浓度的增加,最大吸收波长红移,说明PR-1a中色氨酸残基主要存在于分子内部,且易受温度等因素影响从而导致三维结构的变化.特定浓度的盐酸胍处理后,蛋白呈现中间态.The GST-PR-1a was induced by optimized conditions,and fluorescence spectra were used as the probe to investigate the thermal stability,urea and guanidine hydrochloride degeneration of purified recombinant GST-PR-1a proteins. The fluorescence intensity of PR-1a was reduced slightly when treated at 50 - 80 ℃. However,it was strongly weakened at 90 ℃. With the urea and guanidine hydrochloride treatment,the maximum absorption wavelength occurs red shift along with the increase of the concentration. The results indicate that tryptophan residues are mostly in the interior of PR-1a protein. Factors such as temperature can change its 3D-structure.After specific concentrations of guanidine hydrochloride treatment,protein presents intermediate state.

关 键 词：PR-1a 内源荧光探针 外源荧光探针 热稳定性 尿素 盐酸胍 三维结构 

分 类 号：Q518.4[生物学—生物化学]