荧光相图法研究猪胰腺α-淀粉酶在脲和盐酸胍溶液中的去折叠过程  

Unfolding Procedure of Porcine Pancreatic α-Amylase by Phase Diagram Method of Fluorescence

作　　者：师江波[1] 边六交[1] 董发昕[2] 

机构地区：[1]西北大学生命科学学院基因工程中心,西安710069 [2]西北大学化学系,西安710069

出　　处：《分析化学》2007年第5期707-710,共4页Chinese Journal of Analytical Chemistry

摘　　要：用荧光相图法分别研究了脲和盐酸胍诱导的猪胰腺α-淀粉酶的去折叠过程。实验结果表明,当脲作为变性剂时,无论变性体系中有无还原剂2-巯基乙醇存在,猪胰腺α-淀粉酶的去折叠过程均只出现一个部分折叠中间体,符合“三态模型”;当盐酸胍作为变性剂时,若变性体系中存在还原剂2-巯基乙醇,猪胰腺α-淀粉酶的去折叠过程符合“三态模型”,而若变性体系中不存在还原剂2-巯基乙醇,猪胰腺α-淀粉酶的去折叠过程会出现两个部分折叠中间态,此过程符合“四态模型”。The unfolding procedure of porcine pancreatic α-amylase separately induced by urea and guanidine hydrochloride has been studied by ＂phase diagram＂ method of fluorescence. When porcine pancreatic α-amylase was denatured by urea, in the absence and presence of the reducing agent 2-mercaptoethanol, there exist only one partially folded intermediate during the unfolding procedure from native state to unfolded state, which indicates that the unfolding procedure of α-amylase follows a three-state model at this time. In the presence of reducing agent 2-mercaptoethanol, the denaturation procedure of porcine pancreatic α-amylase induced by guanidine hydrochloride obeys a typical three-state model; while in the absence of 2-mercaptoethanol, the denaturation procedure of porcine pancreatic α-amylase induced by guanidine hydrochloride follows a four-state model and two partially folded intermediates can be detected.

关 键 词：猪胰腺α-淀粉酶 荧光相图法 去折叠过程 脲 盐酸胍 

分 类 号：Q55[生物学—生物化学]