用发光法测定DnaJ类分子伴侣MRJ对虫荧光素酶体外重折叠的促进作用(英文)  

Luminescence Assays for the Luciferase Refolding Facilitated by Human Chaperone DnaJ Homologue MRJ in Vitro

作　　者：刘亚宁[1] 朱美财[1] 刘成刚[1] 赵新华[1] 陈涛[1] 

机构地区：[1]中国人民解放军空军总医院临床实验科,北京100036

出　　处：《发光学报》2003年第2期113-116,共4页Chinese Journal of Luminescence

基　　金：ProjectsupportedbyNationalNaturalScienceFoundationofChina (698710 3 1)

摘　　要：蛋白质的折叠使新翻译合成的多肽变成有生物功能的空间结构 ,因此蛋白质折叠是后基因组时代的重要工作。而在蛋白质折叠过程中 ,必须依靠分子伴侣的相互作用 ,才能保证蛋白质折叠向正确的方向进行。MRJ是一类新发现的DnaJ类分子伴侣 ,已证实它有调节ATP酶活性和阻止多聚谷氨酰胺凝聚的作用。我们用胍变性的虫荧光素酶作为蛋白质恢复功能活性的分子模型 ,用生物发光法研究了MRJ对蛋白质重新折叠的作用 ,发现MRJ确实有促进变性虫荧光素酶重新折叠并恢复其催化活力的功能。而且这种过程需要另一类Hsp6 0和Hsp70的协助 ,而这三种分子伴侣组合在一起 ,功效最好。实验中还发现 ,MRJ对虫荧光素酶恢复的促进作用是与ATP密切相关的。发光分析是研究分子伴侣促进蛋白质折叠过程的一种灵敏、精确和快速的技术手段。A set of specified protein,the molecular chaperones,could bind with other proteins which are not in native state and could promote them to achieve their functional conformations.MRJ (Mouse Related Jprotein) is a kind of newly discovered chaperones and is the homologue of human DnaJ.By using denatured luci ferase as a molecular model to study the protein refolding and renaturation proc esses,the authors incubated the denatured luciferase with three kinds of chaper ones:MRJ,Hsp60,Hsp70,and their different combinations,respectively.Then ,the bioluminescence from luciferinATPMg system,which catalyzed by differe nt ly treated luciferase,was measured.The results showed that all these chaperone s,MRJ,Hsp60 and Hap70,could somewhat increase the luminescence from the syste ms.But only the combination of these three chaperones did have the most signifi cant and most stable effects.The authors also found the whole processes were ob viously ATPdependent.From the experiment the author concluded that (1) As a m o lecular chaperone,MRJ could facilitate the refolding of denatured luciferase; (2) Luminescence assay was a sensitive,accurate and quick method for the chape rone studies.

关 键 词：DnaJ分子伴侣 MRJ 蛋白质折叠 发光分析 测定 虫荧光素酶 体外重新折叠 变性 

分 类 号：Q518.4[生物学—生物化学]