燐光探针法研究不同变性剂对大肠杆菌碱性磷酸酶构象的影响  

Effect of Different Denaturants on Conformation of Escherichia Coli Alkaline Phosphatase by Room Temperature Phosphorescence Probe

作　　者：张海容[1] 晋卫军[1] 刘长松[1] 

机构地区：[1]山西大学化学系

出　　处：《分析化学》2002年第9期1067-1069,共3页Chinese Journal of Analytical Chemistry

基　　金：国家自然科学基金资助课题 (No .2 95 75 2 0 4;2 9875 0 16 )

摘　　要：通过监测大肠杆菌碱性磷酸酶 (AP)的 10 9位色氨酸 (Trp 10 9)室温光 (RTP)的强度与寿命的变化 ,探讨了变性剂酸、盐酸胍及EDTA对AP构象变化的影响。结果表明 :光强度、光寿命与Trp 10 9所处的微环境刚性化程度有密切的关系。尤其是盐酸胍的加入 ,AP经历了典型的 3种变化状态 :从稳定折叠态到中间态 ,最后形成展开态。极少量的EDTA加入 ,导致蛋白质变性、光减弱和光寿命缩短。Effect of different denaturants such as acid, guanidine hydrochloride(Gdn·HCl), ethylene diamine tetraacetic acid (EDTA) on the conformation of Escherichia coli alkaline phosphatase (AP) are explored by determining the room temperature phosphorescence (RTP) intensity and lifetime of tryptophan residues (Trp-109) of AP. The RTP intensity and lifetime are closely associated with the local microenvironmental rigidity of Tp-109. AP has typically undergone three-state changes with the increasing concentration of Gdn·HCl: from stable folded state to intermediate state, and further to final unfolded state. As a trace amount of EDTA is added, which often results in denaturation of protein, the phosphorescence intensity and lifetime are decreased.

关 键 词：Lin光探针法 变性剂 大肠杆菌碱性磷酸酶 室温Lin光 蛋白质 构象 色氨酸 

分 类 号：O657.3[理学—分析化学]