Differential inhibition of endothelial cell proliferation and migration by urokinase subdomains: amino-terminal fragment and kringle domain.
- Author:
Kwang Sei KIM
1
;
Yong Kil HONG
;
Yoon LEE
;
Joo Young SHIN
;
Soo Ik CHANG
;
Soo Il CHUNG
;
Young Ae JOE
Author Information
1. Cancer Research Institute, Catholic Research Institute of Medicial Sciences, The Catholic University of Korea, Seoul 137-701, Korea. youngjoe@catholic.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
angiogenesis inhibitors;
angiostatic proteins;
cell migration inhibition;
endothelial cells;
urokinase-type plasminogen activator
- MeSH:
Animals;
Biosensing Techniques;
Cattle;
Cell Division/drug effects;
Cell Movement/*drug effects;
Cells, Cultured;
Chickens;
Cricetinae;
Endothelial Cells/*cytology/*drug effects;
Humans;
Kinetics;
*Kringles;
Ligands;
Peptide Fragments/*chemistry/genetics/metabolism/*pharmacology;
Protein Binding;
Receptors, Cell Surface/metabolism;
Receptors, Urokinase Plasminogen Activator;
Urokinase-Type Plasminogen Activator/*chemistry/genetics/pharmacology;
Vascular Endothelial Growth Factor A/pharmacology
- From:Experimental & Molecular Medicine
2003;35(6):578-585
- CountryRepublic of Korea
- Language:English
-
Abstract:
The serine protease urokinase-type plasminogen activator (uPA) is implicated in pericellular proteolysis in a variety of physiological and pathological processes including angiogenesis and tumor metastasis. The kringle domain of uPA (UK1) has proven to be an anti-angiogenic molecule with unknown mechanism and amino terminal fragment of uPA (u-ATF) with additional growth factor-like domain can be used for blocking interaction of uPA and uPA receptor. Here, we compared anti-angiogenic activities of these two molecules in vitro and in vivo. The recombinant u-ATF from E. coli and refolded in vitro was found to bind to uPAR with high affinity, whereas E. coli-derived UK1 showed no binding by Biacore analysis. In contrast to UK1 having potent inhibitory effect, u-ATF exhibited low inhibitory effect on bovine capillary endothelial cell growth (ED(50)>320 nM). Furthermore, u-ATF inhibition of VEGF-induced migration of human umbilical vein endothelial cell was far less sensitive (IC(50)= 600 nM) than those observed with UK1, and angiogenesis inhibition was marginal in chorioallantoic membrane. These results suggest that kringle domain alone is sufficient for potent anti- angiogenic activity and additional growth factor-like domain diverts this molecule in undergoing different mechanism such as inhibition of uPA/uPAR interaction rather than undergoing distinct anti- angiogenic mechanism driven by kringle domain.
