还原型牛胰岛素在疏水界面上的折叠

Studieson the refolding of reduced-denatured insulin on a hydrophobic surface

用疏水色谱 ( HPHIC)对还原变性的牛胰岛素在疏水界面上的折叠规律进行了研究 ,对于未还原的胍变和脲变 insulin来说 ,胍变 insulin用 HPHIC法可以完全复性 ,而脲变 insulin只能部分复性。对于还原变性的 insulin,HPHIC对胍变、脲变 insulin只能部分复性。在非氧化型流动相中 ,它们的复性情况较差 ,而采用氧化型流动相 ,有助于二硫键的正确对接 ,可大大提高胍变和脲变 insulin的复性效率。此外 ,还发现有 insulin折叠中间体存在。

The refolding of redcuced- denatured insulin from bovine pancreas on the hydrophobic surface was studied with high performance hydrophobic interaction chromatography ( HPHIC) .It was shown that insulin denatured with 7.0 mol/L guanidine hydrochloride( Gua HCl) can be renatured completely with HPHIC,but urea- unfolded insulin only can berefolded partly.The reduced- denatured insulin also can be refolded partly with HPHIC.In the presence of oxidized glutathione( GSSG) in the mobile phase employed,the disulfide exchange of reduced- denatured insulin can be accelerated and the threedisulfide bridgesof insulin were formed correctly. The resultshowed thatthe refolding yeild ofreduced- denatured insulin with Gua HClor urea should beincreased higher.In addition,the intermediate of urea- unfolded insulin was found and separated with HPHIC.