Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase
文献类型：期刊论文
| 作者 | Gao, Ling1,2; Zhang, Chun1 ; Li, Lingling1; Liang, Lan1; Deng, Xuan1; Wu, Wutong3; Su, Zhiguo2; Yu, Rong1
|
| 刊名 | PROTEIN JOURNAL
 |
| 出版日期 | 2012-04-01 |
| 卷号 | 31期号:4页码:328-336 |
| 关键词 | Bifunctional fusion protein Hirudin-PA C-terminal 12-residue Reteplase Inclusion body Refolding |
| ISSN号 | 1572-3887 |
| 通讯作者 | Yu, R |
| 英文摘要 | To obtain a bifunctional protein simultaneously showing bioactivity of anticoagulant and fibrinolytic for use in the treatment of thrombotic diseases, we constructed a fusion protein (HV12p-rPA) containing C-terminal 12-residue of hirudin-PA (HV12p) and reteplase (rPA). The fusion protein, in which HV12p was linked to rPA via Gly-Gly-Gly, was successfully expressed in an inactive form of inclusion bodies in Escherichia coli. HV12p-rPA was identified by sodium dodecylsulfate-polyacrylamide gel electrophoresis. The expression level of HV12p-rPA was optimized by an orthogonal method and finally enhanced from 12 % to approximate 30 %. We also deeply investigated the condition of renaturation of HV12p-rPA, and the inactive protein was partly renatured through various conditions. The refolding efficacy of HV12p-rPA estimated by the recovery of fibrinolytic activity varied from 0.03 % to 16.6 % and the anticoagulant activity fluctuated in the range from 41 to 2,297 ATU/mg. Bioassays indicated that the resulted fusion protein, as expected, exhibited both fibrinolytic and anticoagulant activities. These works laid a foundation for further characterization of HV12p-rPA. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biochemistry & Molecular Biology |
| 研究领域[WOS] | Biochemistry & Molecular Biology |
| 关键词[WOS] | TISSUE-PLASMINOGEN ACTIVATOR ; ACUTE MYOCARDIAL-INFARCTION ; ACUTE ISCHEMIC-STROKE ; INCLUSION-BODY PROTEINS ; THROMBOLYTIC THERAPY ; THROMBIN INHIBITION ; CANINE MODEL ; HEPARIN ; STREPTOKINASE ; ANTICOAGULANT |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000303456800007 |
| 公开日期 | 2013-10-18 |
| 版本 | 出版稿 |
| 源URL | [http://ir.ipe.ac.cn/handle/122111/3824]  |
| 专题 | 过程工程研究所_生化工程国家重点实验室 |
| 作者单位 | 1.Sichuan Univ, Key Lab Drug Targeting & Drug Delivery Syst, Minist Educ, W China Sch Pharm, Chengdu 610041, Peoples R China 2.Chinese Acad Sci, State Key Lab Biochem Engn, Inst Proc Engn, Beijing 100080, Peoples R China 3.China Pharmaceut Univ, Sch Life Sci & Technol, Nanjing 210009, Peoples R China |
| 推荐引用方式 GB/T 7714 | Gao, Ling,Zhang, Chun,Li, Lingling,et al. Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase[J]. PROTEIN JOURNAL,2012,31(4):328-336. |
| APA | Gao, Ling.,Zhang, Chun.,Li, Lingling.,Liang, Lan.,Deng, Xuan.,...&Yu, Rong.(2012).Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase.PROTEIN JOURNAL,31(4),328-336. |
| MLA | Gao, Ling,et al."Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase".PROTEIN JOURNAL 31.4(2012):328-336. |
入库方式： OAI收割
来源：过程工程研究所
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