Studies on the refolding of the reduced-denatured insulin with size exclusion chromatography  

作　　者：BAI Quan KONG Yu DONG Cuihua GENG Xindu 

机构地区：[1]Institute of Modern Separation Science,Shaanxi Key Laboratory of Modern Separation Science,Northwest University,Xi'an 710069,China

出　　处：《Science China Chemistry》2005年第z1期55-59,共5页中国科学(化学英文版)

基　　金：supported by the National Natural Science Foundation of China(Grant Nos.29675017&39880003);the Foundation of the Young Teacher in the University of Ministry of Education,China(Grant No.DF00308);the Natural Science Foundation in Shaanxi Province(Grant No.2001H02);the Foundation of the Committee of Education in Shaanxi Province(Grant No.99JK101).

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(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).

关 键 词：protein folding size exclusion chromatography reduced-denaturation disulfide bonding INSULIN 

分 类 号：N[自然科学总论]