PMID- 23751561
OWN - NLM
STAT- MEDLINE
DCOM- 20140407
LR  - 20190923
IS  - 1738-8872 (Electronic)
IS  - 1017-7825 (Linking)
VI  - 23
IP  - 9
DP  - 2013 Sep 28
TI  - Expression, purification, and biological characterization of the amino-terminal 
      fragment of urokinase in Pichia pastoris.
PG  - 1197-205
AB  - Urokinase (uPA) and its receptor (uPAR) play an important role in tumor growth 
      and metastasis. Targeting the excessive activation of this system as well as the 
      proliferation of the tumor vascular endothelial cell would be expected to prevent 
      tumor neovasculature and halt the tumor development. In this regard, the 
      amino-terminal fragment (ATF) of urokinase has been confirmed as effective to 
      inhibit the proliferation, migration, and invasiveness of cancer cells via 
      interrupting the interaction of uPA and uPAR. Previous studies indicated that ATF 
      expressed in Escherichia coli was mainly contained in inclusion bodies and also 
      lacked posttranslational modifications. In this study, the biologically active 
      and soluble ATF was cloned and expressed in Pichia pastoris. The recombinant 
      protein was purified to be homogenous and confirmed to be biologically active. 
      The yield of the active ATF was about 30 mg/l of the P. pastoris culture medium. 
      The recombinant ATF (rATF) could efficiently inhibit angiogenesis, endothelial 
      cell migration, and tumor cell invasion in vitro. Furthermore, it could inhibit 
      in vivo xenograft tumor growth and prolong the survival of tumor-bearing mice 
      significantly by competing with uPA for binding to cell surfaces. Therefore, P. 
      pastoris is a highly efficient and cost-effective expression system for 
      large-scale production of biologically active rATFs for potential therapeutic 
      application.
FAU - Li, Jianping
AU  - Li J
AD  - The State Key Laboratory of Pharmaceutical Biotechnology, Nanjing University, 
      Nanjing 210093 Jiangsu, P.R. China.
FAU - Lin, Yuli
AU  - Lin Y
FAU - Zhuang, Hongqin
AU  - Zhuang H
FAU - Hua, Zi-Chun
AU  - Hua ZC
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - Korea (South)
TA  - J Microbiol Biotechnol
JT  - Journal of microbiology and biotechnology
JID - 9431852
RN  - 0 (Angiogenesis Inhibitors)
RN  - 0 (Recombinant Proteins)
RN  - EC 3.4.21.73 (Urokinase-Type Plasminogen Activator)
SB  - IM
MH  - Amino Acid Motifs
MH  - Angiogenesis Inhibitors/chemistry/genetics/*isolation & 
      purification/*pharmacology
MH  - Animals
MH  - Cell Line, Tumor
MH  - Cell Movement/drug effects
MH  - Chick Embryo
MH  - Chorioallantoic Membrane/blood supply/drug effects
MH  - *Gene Expression
MH  - Humans
MH  - Mice
MH  - Mice, Nude
MH  - Neovascularization, Pathologic/drug therapy
MH  - Pichia/*genetics/metabolism
MH  - Recombinant Proteins/chemistry/genetics/isolation & purification/pharmacology
MH  - Urokinase-Type Plasminogen Activator/chemistry/genetics/*isolation & 
      purification/*pharmacology
EDAT- 2013/06/12 06:00
MHDA- 2014/04/08 06:00
CRDT- 2013/06/12 06:00
PHST- 2013/06/12 06:00 [entrez]
PHST- 2013/06/12 06:00 [pubmed]
PHST- 2014/04/08 06:00 [medline]
AID - 10.4014/jmb.1305.05004 [pii]
AID - 10.4014/jmb.1305.05004 [doi]
PST - ppublish
SO  - J Microbiol Biotechnol. 2013 Sep 28;23(9):1197-205. doi: 10.4014/jmb.1305.05004.