| Studies on the refolding of the reduced-denatured insulin with size exclusion chromatography |
| |
| 作者姓名： | BAI Quan KONG Yu DONG Cuihua & GENG Xindu Institute of Modern Separation Science Shaanxi Key Laboratory of Modern Separation Science Northwest University Xi'an China |
| |
| 作者单位： | BAI Quan，KONG Yu，DONG Cuihua & GENG Xindu Institute of Modern Separation Science，Shaanxi Key Laboratory of Modern Separation Science，Northwest University，Xi'an 710069，China |
| |
| 基金项目： | China，陕西省自然科学基金，the Foundation of the Committee of Education in Shaanxi Province |
| |
| 摘 要： | The refolding of the reduced-denatured insulin from bovine pancreas was investigated with the size exclusion chromatography (SEC). It was shown that the reduced-denatured insulin originally denatured with 7.0 mol·L-1 guanidine hydrochloride (GuHCI) or 8.0 mol·L-1 urea could not be refolded with a non-oxidized mobile phase. Although the oxidized and reduced glutathione (GSSG and GSH) were employed in the oxidized mobile phase, the reduced-denatured insulin still could not be renatured. However, in the presence of 2.0 mol·L-1 urea in the oxidized mobile phase employed, the reduced-denatured insulin can be refolded with SEC, and the aggregation of denatured insulin can be diminished by urea. In addition, the disul-fide exchange of reduced-denatured insulin also can be accelerated with GSSG/GSH in the oxidized mobile phase. The three disulfide bridges of insulin were formed correctly and the reduced-unfolded insulin can be renatured completely. The results were further tested with re-versed-phase liquid chromatography (RPLC) and hydrophobic interaction chromatography (HIC).
|
Studies on the refolding of the reduced-denatured insulin with size exclusion chromatography |
| |
| BAI Quan,KONG Yu,DONG Cuihua & GENG Xindu Institute of Modern Separation Science,Shaanxi Key Laboratory of Modern Separation Science,Northwest University,Xi''''an ,China.Studies on the refolding of the reduced-denatured insulin with size exclusion chromatography[J].Science in China(Chemistry),2005,48(Z1). |
| |
| Authors: | BAI Quan KONG Yu DONG Cuihua GENG Xindu |
| |
| Institution: | Institute of Modern Separation Science, Shaanxi Key Laboratory of Modern Separation Science, Northwest University, Xi'an 710069, China |
| |
| Abstract: | The refolding of the reduced-denatured insulin from bovine pancreas was investigated with the size exclusion chromatography (SEC). It was shown that the reduced-denatured insulin originally denatured with 7.0 mol·L-1 guanidine hydrochloride (GuHCl) or 8.0 mol·L-1 urea could not be refolded with a non-oxidized mobile phase. Although the oxidized and reduced glutathione (GSSG and GSH) were employed in the oxidized mobile phase, the reduced-denatured insulin still could not be renatured. However, in the presence of 2.0 mol·L-1 urea in the oxidized mobile phase employed, the reduced-denatured insulin can be refolded with SEC, and the aggregation of denatured insulin can be diminished by urea. In addition, the disulfide exchange of reduced-denatured insulin also can be accelerated with GSSG/GSH in the oxidized mobile phase. The three disulfide bridges of insulin were formed correctly and the reduced-unfolded insulin can be renatured completely. The results were further tested with reversed-phase liquid chromatography (RPLC) and hydrophobic interaction chromatography (HIC). |
| |
| Keywords: | protein folding size exclusion chromatography reduced-denaturation disulfide bonding insulin |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
