Expression and characterization of recombinant human micro-plasminogen

详细信息    查看全文

• 作者：Zhifeng Ma ; Wei Lu ; Sheng Wu ; Junyong Chen ; Ziyong Sun and Jian-Ning Liu
• 关键词：Expression ; Micro ; plasminogen ; Prourokinase ; Purification ; Urokinase
• 刊名：Biotechnology Letters
• 出版年：2007
• 出版时间：April, 2007
• 年：2007
• 卷：29
• 期：4
• 页码：517-523
• 全文大小：201 KB
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Microbiology
  Biotechnology
  Applied Microbiology
  Biochemistry
  
• 出版者：Springer Netherlands
• ISSN：1573-6776

文摘

Micro-plasminogen (μPlg) gene amplified from human liver cells by reverse transcription PCR was inserted into expression vector pET-28a (pET-28a/μPlg) and transformed into E. coli strain BL21(DE3). Recombinant human micro-plasminogen (rh-μPlg) was over-expressed as inclusion bodies when induced with IPTG. After renaturation and purification, 16 mg rh-μPlg/l was obtained with a homogeneity of 95% (w/w). Pro-urokinase (proUK)-induced rh-μPlg activation was significantly faster than when Glu-plasminogen was the substrate. The catalytic efficiency of urokinase (UK) activation of rh-μPlg was twice that of Glu-plasminogen. While recombinant human micro-plasmin (rh-μPlm) and Lys-plasmin had a similar amidolytic activity against a small substrate, d-valyl-l-leucyllysine-p-nitroaniline dihydrochloride, Lys-plasmin activated proUK with a catalytic efficiency about fourfold greater than did rh-μPlm. These results suggested that the kringle 1–5 domain of plasminogen and plasmin may modify both UK activation of plasminogen and plasmin activation of proUK, respectively.