Anti-angiogenic action of plasma hyaluronan binding protein in human umbilical vein endothelial cells.
The kringle domain is a triple loop structure present in angiostatin and endostatin. The disulfide bond-linked kringle architectures have been known to be essential for anti-angiogenic activity. Plasma hyaluronan binding protein (PHBP) is a novel serine protease which consists of three epidermal growth factor (EGF) domains, a kringle domain, and a serine protease domain. PHBP can be cleaved autocatalytically to generate activity and is highly expressed in the human blood and liver. To determine the anti-angiogenic activities of PHBP, we purified recombinant mouse PHBP from stable cell line overexpressing PHBP and used protein in vivo and in vitro angiogenesis assays. We found that recombinant PHBP inhibits not only angiogenesis in vivo in chorioallantoic membrane (CAM) assay but also the basic fibroblast growth factor (bFGF)-induced proliferation, invasion and tube formation of human umbilical vein endothelial cells (HUVECs) in a dose-dependant manner. Moreover, we found that the kringle domain of PHBP was essential for the anti-angiogenic action of PHBP by the deletion mutants. These findings unravel a new function of PHBP as an inhibitor of the proangiogenic phenotype of vascular endothelial cells and demonstrate that the kringle domain of PHBP might be a potent novel inhibitor of activated endothelial cells in vitro and in vivo.
Jeon
Ji Won
JW
Department of Molecular Biology, Pusan National University, Busan 609-735, South Korea.
Song
Hyun Seok
HS
Moon
Eun-Joung
EJ
Park
Shi-Young
SY
Son
Myung Jin
MJ
Jung
Seung Youn
SY
Kim
Ji Tae
JT
Nam
Do-Hyun
DH
Choi-Miura
Nam-Ho
NH
Kim
Kyu-Won
KW
Kim
Yung-Jin
YJ
D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D000818:Animals; D002465:Cell Movement; D049109:Cell Proliferation; D002478:Cells, Cultured; D002642:Chick Embryo; D049033:Chorioallantoic Membrane / Q000098:blood supply / Q000187:drug effects; D003094:Collagen; D004305:Dose-Response Relationship, Drug; D004338:Drug Combinations; D042783:Endothelial Cells / Q000166:cytology / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology; D006801:Humans; D018082:Kringles / Q000235:genetics; D007797:Laminin; D009154:Mutation; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D011509:Proteoglycans; D011994:Recombinant Proteins / Q000737:chemistry / Q000494:pharmacology; D012697:Serine Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000494:pharmacology*; D014471:Umbilical Veins / Q000166:cytology / Q000187:drug effects
D020533:Angiogenesis Inhibitors; D004338:Drug Combinations; D007797:Laminin; D011509:Proteoglycans; D011994:Recombinant Proteins; D016222:Fibroblast Growth Factor 2; C059485:matrigel; D003094:Collagen; C499020:Habp2 protein, mouse; D012697:Serine Endopeptidases