Differential inhibition of endothelial cell proliferation and migration by urokinase subdomains: amino-terminal fragment and kringle domain.

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.

• Authors: Kwang Sei Kim, Yong-Kil Hong, Yoon Lee, Joo-Young Shin, Soo-Ik Chang, Soo Il Chung, Young Ae Joe
• Journal: Experimental & molecular medicine (Exp Mol Med) Vol. 35 Issue 6 Pg. 578-85 (Dec 31 2003) ISSN: 1226-3613 [Print] United States
• PMID: 14749538 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Ligands
• PLAUR protein, human
• Peptide Fragments
• Receptors, Cell Surface
• Receptors, Urokinase Plasminogen Activator
• Vascular Endothelial Growth Factor A
• Urokinase-Type Plasminogen Activator

Topics

• 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)