作　　者： ; ; ; ; ; ;

机构地区： 南京理工大学化工学院

出　　处： 《中国生物工程杂志》 2009年第8期26-32,共7页

摘　　要： 为考察一种对pH较为敏感的多聚化合物聚丙烯酸树脂Eudragit S-100对人转化生长因子β1(transforming growth factor,TGF-β1)复性的作用,将以包涵体形式存在TGF-β1进行变性,并将变性蛋白直接加入到含有不同浓度Eudragit的蛋白复性缓冲液中。采用MTT法、荧光分光光度法、圆二色谱以及高效液相色谱法等方法来比较分析不同浓度Eudragit S-100对变性TGF-β1的复性促进作用。实验结果表明,在Eudragit S-100作用下TGF-β1的复性产率比普通稀释复性法显著增高,且最高达到53%,研究还表明Eudragit S-100的促进蛋白复性的作用是基于Eudragit S-100与TGF-β1发生了特异性的离子结合反应。通过这一反应,Eudragit S-100遮蔽了蛋白多肽间的疏水基团,有效地抑制了蛋白的聚集进而发挥其促复性功能。 If Eudragit S-100, a pH-responsive polymer can enhance the refolding level of TGF-β1 （transforwing growth factor,TGF-β1 ）. The refolding of TGF-β1 was performed by directly diluting denatured TGF-β1 into a refolding buffer containing different concentration of Eudragit. The ability of Eudragit S-100 to enhance protein refolding level was investigated using MTT assay, reverse phase HPLC, fluorescence emission spectroscopy and circular dichroism spectroscopy. The addition of Eudragit S-100 in the refolding buffer significantly increased TGF-β1 refolding yield to 53%, when dilution refolding was conducted at 0.5 mg/ml TGF -β1. This shows the evidence of an electrostatic interaction between oppositely charged TGF-β1 and the Eudragit polymer during refolding. This ionic complex of Eudragit and TGF-β1 appears to shield exposed hydrophobic residues of the lysozyme refolding intermediates, thus minimizing hydrophobicdriven aggregation of the molecules.

关 键 词： 聚丙烯酸树脂 人转化生长因子 蛋白复性

领　　域： [生物学]