In vitro refolding of human proinsulin - Kinetic intermediates, putative disulfide-forming pathway, folding initiation site, and potential role of C-peptide in folding process
文献类型：期刊论文
| 作者 | Qiao, ZS; Min, CY; Hua, QX; Weiss, MA; Feng, YM |
| 刊名 | JOURNAL OF BIOLOGICAL CHEMISTRY
 |
| 出版日期 | 2003 |
| 卷号 | 278期号:20页码:17800-17809 |
| 通讯作者 | Feng, YM (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, 320 Yue Yang Rd, Shanghai 200031, Peoples R China., |
| 英文摘要 | Human insulin is a double-chain peptide that is synthesized in vivo as a single-chain human proinsulin (HPI). We have investigated the disulfide-forming pathway of a single-chain porcine insulin precursor (PIP). Here we further studied the folding pathway of HPI in vitro. While the oxidized refolding process of HPI was quenched, four obvious intermediates (namely P1, P2, P3, and P4, respectively) with three disulfide bridges were isolated and characterized. Contrary to the folding pathway of PIP, no intermediates with one- or two-disulfide bonds could be captured under different refolding conditions. CD analysis showed that P1, P2, and P3 retained partially structural conformations, whereas P4 contained little secondary structure. Based on the time-dependent distribution, disulfide pair analysis, and disulfide-reshuffling process of the intermediates, we have proposed that the folding pathway of HPI is significantly different from that of PIP. These differences reveal that the C-peptide not only facilitates the folding of HPI but also governs its kinetic folding pathway of HPI. Detailed analysis of the molecular folding process reveals that there are some similar folding mechanisms between PIP and HPI. These similarities imply that the initiation site for the folding of PIP/HPI may reside in the central alpha-helix of the B-chain. The formation of disulfide A20-B19 may guide the transfer of the folding information from the B-chain template to the unstructured A-chain. Furthermore, the implications of this in vitro refolding study on the in vivo folding process of HPI have been discussed. |
| 学科主题 | Biochemistry & Molecular Biology |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | GROWTH-FACTOR-I ; PANCREATIC TRYPSIN-INHIBITOR ; ENGINEERED INSULIN MONOMER ; STRUCTURAL INFORMATION ; MASS-SPECTROMETRY ; NATIVE MOLECULE ; RIBONUCLEASE-A ; SMALL PROTEINS ; IGF-I ; BPTI |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000182838300028 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2383]  |
| 专题 | 上海生化细胞研究所_上海生科院生化细胞研究所 |
| 推荐引用方式 GB/T 7714 | Qiao, ZS,Min, CY,Hua, QX,et al. In vitro refolding of human proinsulin - Kinetic intermediates, putative disulfide-forming pathway, folding initiation site, and potential role of C-peptide in folding process[J]. JOURNAL OF BIOLOGICAL CHEMISTRY,2003,278(20):17800-17809. |
| APA | Qiao, ZS,Min, CY,Hua, QX,Weiss, MA,&Feng, YM.(2003).In vitro refolding of human proinsulin - Kinetic intermediates, putative disulfide-forming pathway, folding initiation site, and potential role of C-peptide in folding process.JOURNAL OF BIOLOGICAL CHEMISTRY,278(20),17800-17809. |
| MLA | Qiao, ZS,et al."In vitro refolding of human proinsulin - Kinetic intermediates, putative disulfide-forming pathway, folding initiation site, and potential role of C-peptide in folding process".JOURNAL OF BIOLOGICAL CHEMISTRY 278.20(2003):17800-17809. |
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