感光受体外周蛋白结合蛋白对虫荧光素酶体外折叠的促进作用  

Improvement of luciferase refolding in vitro by peripherin-binding protein of photoreceptor cell

作　　者：刘成刚[1] 朱美财[1] 占志[1] 王荫静[1] 李文[1] 刘亚宁[1] 

机构地区：[1]空军总医院临床分子生物学中心,北京100036

出　　处：《基础医学与临床》2002年第6期570-573,共4页Basic and Clinical Medicine

基　　金：国家自然科学基金 (6 98710 31) ;总后医药卫生基金 (0 1MA0 6 0 )

摘　　要：探讨感光受体外周蛋白结合蛋白 (PBP)在体外是否有促进变性蛋白质复性的作用。虫荧光素酶用盐酸胍变性 ,然后在PBP、HSP70、HSP6 0存在下进行体外复性 ,用虫荧光素酶分析系统检测该酶的复性程度。结果显示PBP对虫荧光素酶的复性能力很低 ,当与HSP70同时作用时 ,酶活力明显高于PBP或HSP70组 ,而PBP与HSP6 0组合对酶的复性能力较HSP6 0组未见有明显差异 ;当PBP ,HSP70 ,HSP6 0三者同时存在时 ,酶活力恢复率最高。去除复性缓冲液中的ATP及其再生系统 ,酶活力的复性率明显下降。结果表明PBP具有协助HSP70 。The possibility of peripherin binding protein (PBP) to improve the refolding of denatured luciferase in vitro was investigated. Luciferase was denatured with 6mol/L guanidine hydrochlori and then added into refolding buffer containing different molecular chaperone including PBP, HSP70, HSP60 and their combinations. Sample was analyzed with luciferase assay solution. The enzyme activity raised a little after PBP or HSP70 being added, however, the recovery of enzyme activity had significantly raised when the reaction was conducted with PBP plus HSP70. However, PBP+HSP60 and HSP70+HSP60 have no such effect compare to HSP60. PBP+HSP70+HSP60 has the most refolding yield of denatured enzyme in all molecular chaperones. The refolding was markedly decreased after ATP being depleted . It is concluded that PBP cooperates with HSP70 and to improve productive folding of chemically denatured luciferase in ATP dependent in vitro .

关 键 词：感光受体外周蛋白结合蛋白 热休克蛋白 虫荧光素酶 复性 

分 类 号：R346[医药卫生—基础医学]