Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase

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	Construction, Expression and Refolding of a Bifunctional Fusion Protein Consisting of C-Terminal 12-Residue of Hirudin-PA and Reteplase
	Gao, Ling 1,2; Zhang, Chun1 ; Li, Lingling 1; Liang, Lan 1; Deng, Xuan 1; Wu, Wutong 3; Su, Zhiguo 2; Yu, Rong 1
刊名	PROTEIN JOURNAL
	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
内容类型	期刊论文
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.