Purification, cDNA cloning, and recombinant expression of chymotrypsin C from porcine pancreas
文献类型：期刊论文
| 作者 | Wang, HB; Yuan, DD; Xu, R; Chi, CW |
| 刊名 | ACTA BIOCHIMICA ET BIOPHYSICA SINICA
 |
| 出版日期 | 2011 |
| 卷号 | 43期号:7页码:568-575 |
| 关键词 | porcine pancreas sequence determination gene cloning gene expression |
| 通讯作者 | Chi, CW (reprint author), Chinese Acad Sci, Shanghai Inst Biol Sci, Inst Biochem & Cell Biol, State Key Lab Mol Biol, Shanghai 200031, Peoples R China.,zwqi@sibs.ac.cn |
| 英文摘要 | Chymotrypsin C is a bifunctional secretory-type serine protease in pancreas; besides proteolytical activity, it also exhibits a calcium-decreasing activity in serum. In this study, we purified activated chymotrypsin C from porcine pancreas, and identified its three active forms. Active chymotrypsin C was found to be different in the length of its 13-residue activation peptide due to carboxydipeptidase (present in the pancreas) degradation or autolysis of the activated chymotrypsin C itself, resulting in the removal of several C-terminus residues from the activation peptide. After limited chymotrypsin C cleavage with endopeptidase Lys C, several purified peptides were partially sequenced, and the entire cDNA sequence for porcine chymotrypsin C was cloned. Recombinant chymotrypsinogen C was successfully expressed in Escherichia coli cells as inclusion bodies. After refolding and activation with trypsin, the comparison of the recombinant chymotrypsin C with the natural form showed that their proteolytic and calcium-decreasing activities were at the same level. The successful expression of chymotrypsin C gene paves the way to further mutagenic structure-2 function studies. |
| 学科主题 | Biochemistry & Molecular Biology; Biophysics |
| 类目[WOS] | Biochemistry & Molecular Biology ; Biophysics |
| 关键词[WOS] | CALCIUM-DECREASING FACTOR ; FACTOR CALDECRIN ; MOLECULAR-CLONING ; CRYSTAL-STRUCTURE ; CATHEPSIN-B ; ACTIVATION ; SPECIFICITY ; TRYPSINOGEN ; SUPPRESSION ; SEQUENCE |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000292337100009 |
| 版本 | 出版稿 |
| 源URL | [http://202.127.25.143/handle/331003/832]  |
| 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 | Wang, HB,Yuan, DD,Xu, R,et al. Purification, cDNA cloning, and recombinant expression of chymotrypsin C from porcine pancreas[J]. ACTA BIOCHIMICA ET BIOPHYSICA SINICA,2011,43(7):568-575. |
| APA | Wang, HB,Yuan, DD,Xu, R,&Chi, CW.(2011).Purification, cDNA cloning, and recombinant expression of chymotrypsin C from porcine pancreas.ACTA BIOCHIMICA ET BIOPHYSICA SINICA,43(7),568-575. |
| MLA | Wang, HB,et al."Purification, cDNA cloning, and recombinant expression of chymotrypsin C from porcine pancreas".ACTA BIOCHIMICA ET BIOPHYSICA SINICA 43.7(2011):568-575. |
入库方式： OAI收割
浏览0
下载0
收藏0
其他版本
除非特别说明，本系统中所有内容都受版权保护，并保留所有权利。