Domain 5 of high molecular weight kininogen (kininostatin) down-regulates endothelial cell proliferation and migration and inhibits angiogenesis

被引:154
作者
Colman, RW
Jameson, BA
Lin, YZ
Johnson, D
Mousa, SA
机构
[1] Temple Univ, Sol Sherry Thrombosis Res Ctr, Sch Med, Philadelphia, PA 19140 USA
[2] MCP Hahnemann Med Sch, Ctr Neurovirol, Philadelphia, PA USA
[3] Dupont Merck Pharmaceut Co, Div Cardiovasc, Wilmington, DE 19880 USA
关键词
D O I
10.1182/blood.V95.2.543
中图分类号
R5 [内科学];
学科分类号
1002 ; 100201 ;
摘要
We have demonstrated that high molecular weight kininogen (HK) binds specifically on endothelial cells to domain 2/3 of the urokinase receptor (uPAR). Inhibition by vitronectin suggests that kallikrein-cleaved HK (HKa) is antiadhesive. Plasma kallikrein bound to HK cleaves prourokinase to urokinase, initiating cell-associated fibrinolysis. We postulated that HK cell binding,domains would inhibit angiogenesis. We found that recombinant domain 5 (D5) inhibited endothelial cell migration toward vitronectin 85% at 0.27 mu M with an IC50 (concentration to yield 50% inhibition) = 0.12 mu M A D5 peptide, G486-K502, showed an IC50 = 0.2 mu M, but a 25-mer peptide from a D3 cell binding domain only inhibited migration 10% at 139 mu M (IC50 > 50 mu M). D6 exhibited weaker inhibitory activity (IC50 = 0.50 mu M) D5 also potently inhibited endothelial cell proliferation with an IC50 = 30 nM, while D3 and D6 were inactive, Using deletion mutants of D5, we localized the smallest region for full activity to H441-D474. To further map the active region, we created a molecular homology model of D5 and designed a series of peptides displaying surface loops. Peptide 440-155 was the most potent (IC50 = 100 nM) in inhibiting proliferation but did not inhibit migration. D5 inhibited angiogenesis stimulated by fibroblast growth factor FGF2 (97%) in a chicken chorioallantoic membrane assay at 270 nM, and peptide 400-455 was also inhibitory (79%), HK D5 (for which we suggest the designation, "kininostatin") is a potent inhibitor of endothelial cell migration and proliferation in vitro and of angiogenesis in vivo. (Blood, 2000;95:543-550) (C) 2000 by The American Society of Hematology.
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页码:543 / 550
页数:8
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