HPHIC固定相对脲变α-糜蛋白酶复性的影响  

Influence of high-performance hydrophobic interaction chromatographic stationary phase on refolding of urea-denatured α-Chymotrypsin

作　　者：刘振岭[1,2] 柯从玉[2] 耿信笃[2] 

机构地区：[1]新乡医学院化学教研室,新乡453003 [2]西北大学现代分离科学研究所,西安710069

出　　处：《分析试验室》2009年第5期34-37,共4页Chinese Journal of Analysis Laboratory

基　　金：国家自然科学基金(20175016)项目资助

摘　　要：依据计量置换保留理论所得到的参数lgI,来测定不同构象态α-糜蛋白酶(α-Chy)在两种不同高效疏水相互作用色谱(HPHIC)固定相表面的折叠自由能,发现脲变α-Chy在HPHIC固定相表面获取的折叠自由能比溶液中的高很多,不同HPHIC固定相表面为脲变α-Chy提供不同的折叠自由能,且都随变性剂脲浓度的增大而增大;通过对不同HPHIC色谱柱后复性α-Chy的比活测定,还发现脲变α-Chy的复性效率与其从固定相表面的折叠自由能有关,同一构象的α-Chy从固定相表面得到的折叠自由能越高越有利于其折叠成天然蛋白质。The folding free energy of α-chymotrypsin（α-Chy） molecules exist in different molecular conformations on the different surface of stationary phase of high-performance hydrophobic interaction chromatography （HPHIC） could be calculated with the parameters logI values obtained according to stoichiometric displacement theory for retention. It can be found that the urea-denatured α-Chy can take much higher energy on surface of stationary phase of HPHIC than that in aqueous solution, and the different surfaces of stationary phase of HPHIC provide different folding free energies for the urea-denatured α-Chy and the folding free energy of the α-Chy increases with increases in urea concentration. It can also be seen that the efficiencies of refolding of urea-denatured α-Chy are related to the folding free energy on surface of stationary phase of HPHIC by the determination of activities of protein refolding passing through the HIC column. The larger the folding free energy of the same molecular conformation α-Chy takes from the interface of stationary phase of HIC, the higher the possibility of denatured protein refolding to its natural state.

关 键 词：蛋白折叠 疏水相互作用色谱 计量置换理论 Α-糜蛋白酶 折叠自由能 

分 类 号：O558.1[理学—热学与物质分子运动论]