Angiostatin and plasminogen share binding to endothelial cell surface actin.
Previous studies from this laboratory have demonstrated that plasminogen binds to endothelial cell surface-associated actin via its kringles in a dose-dependent and specific manner. The purpose of this study was to determine whether angiostatin, a proteolytic fragment of plasminogen, shares binding properties with plasminogen. Our results indicated that like plasminogen, angiostatin bound to actin in a time-, concentration-, and kringle-dependent manner. Furthermore, this binding was significantly inhibited by excess plasminogen, suggesting that both proteins shared binding motifs on the actin molecule. Fluorescence studies demonstrated that angiostatin bound to intact endothelial cells through its kringles, and this binding was also inhibited by plasminogen but not by unrelated proteins. Ligand blot analyses on endothelial cell lysates indicated that angiostatin interacted with a 42 kDa protein, which was identified as actin. Furthermore, an anti-actin antibody inhibited binding of angiostatin to endothelial cells by approximately 25%. These results suggest that angiostatin and plasminogen share binding to endothelial cell surface actin and, therefore, that angiostatin has the potential to inhibit plasmin-dependent processes such as cell migration-movement.
Dudani
A K
AK
Centre for Biologics Research, Biologics and Genetic Theapies Directorate, Health Canada, Sir Fredrick Banting Research Centre, Ottawa, ON K1A 0L2, Canada. anil_dudani@hc-sc.gc.ca
Ben-Tchavtchavadze
M
M
Porter
S
S
Tackaberry
E
E
D000199:Actins / Q000276:immunology / Q000378:metabolism*; D043168:Angiostatins / Q000378:metabolism*; D000818:Animals; D002462:Cell Membrane / Q000378:metabolism*; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology* / Q000378:metabolism*; D006801:Humans; D018082:Kringles; D008856:Microscopy, Fluorescence; D010958:Plasminogen / Q000378:metabolism*; D011485:Protein Binding; D014471:Umbilical Veins / Q000166:cytology
D000199:Actins; D043168:Angiostatins; D010958:Plasminogen