Unfolding of human proinsulin - Intermediates and possible role of its C-peptide in folding/unfolding
文献类型：期刊论文
| 作者 | Min, CY; Qiao, ZS; Feng, YM |
| 刊名 | EUROPEAN JOURNAL OF BIOCHEMISTRY
 |
| 出版日期 | 2004 |
| 卷号 | 271期号:9页码:1737-1747 |
| 关键词 | C-peptide disulfide scrambling disulfide stability human proinsulin unfolding |
| 通讯作者 | Feng, YM (reprint author), Chinese Acad Sci, Shanghai Inst Biochem, 320 Yue Yang Rd, Shanghai 200031, Peoples R China.,fengym@sunm.shcnc.ac.cn |
| 英文摘要 | We have investigated the in vitro refolding process of human proinsulin (HPI) and an artificial mini-C derivative of HPI (porcine insulin precursor, PIP), and found that they have significantly different disulfide-formation pathways. HPI and PIP differ in their amino acid sequences due to the presence of the C-peptide linker found in HPI, therefore suggesting that the C-peptide linker may be responsible for the observed difference in folding behaviour. However, the manner in which the C-peptide contributes to this difference is still unknown. We have used both the disulfide scrambling method and a redox-equilibrium assay to assess the stability of the disulfide bridges. The results show that disulfide reshuffling is easier to induce in HPI than in PIP by the addition of thiol reagent. Thus, the C-peptide may affect the unique folding pathway of HPI by allowing the disulfide bonds of HPI to be easily accessible. The detailed processes of HPI unfolding by reduction of its disulfide bonds and by disulfide scrambling methods were also investigated. In the reductive unfolding process no accumulation of intermediates was detected. In the process of unfolding by disulfide scrambling, HPI gradually rearranged its disulfide bonds to form three major isomers G1, G2 and G3. The most abundant isomer, G1, contains the B7-B19 disulfide bridge. Based on far-UV CD spectra, native gel analysis and cleavage by endoproteinase V8, the G1 isomer has been shown to resemble the intermediate P4 found in the refolding process of HPI. Finally, the major isomer G1 is allowed to refold to native protein HPI by disulfide rearrangement, which indicates that a similar molecular mechanism may exist for the unfolding and refolding process of HPI. |
| 学科主题 | Biochemistry & Molecular Biology |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | PANCREATIC TRYPSIN-INHIBITOR ; DISULFIDE-FORMING PATHWAY ; CARBOXYPEPTIDASE INHIBITOR ; STRUCTURAL INFORMATION ; ALPHA-LACTALBUMIN ; INSULIN PRECURSOR ; NATIVE MOLECULE ; RIBONUCLEASE-A ; IN-VITRO ; PROTEIN |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000220886100016 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/2130]  |
| 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 | Min, CY,Qiao, ZS,Feng, YM. Unfolding of human proinsulin - Intermediates and possible role of its C-peptide in folding/unfolding[J]. EUROPEAN JOURNAL OF BIOCHEMISTRY,2004,271(9):1737-1747. |
| APA | Min, CY,Qiao, ZS,&Feng, YM.(2004).Unfolding of human proinsulin - Intermediates and possible role of its C-peptide in folding/unfolding.EUROPEAN JOURNAL OF BIOCHEMISTRY,271(9),1737-1747. |
| MLA | Min, CY,et al."Unfolding of human proinsulin - Intermediates and possible role of its C-peptide in folding/unfolding".EUROPEAN JOURNAL OF BIOCHEMISTRY 271.9(2004):1737-1747. |
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