The binding of factor IXa to cultured bovine aortic endothelial cells

David Stern, P. P. Nawroth, W. Kisiel, G. Vehar, C. T. Esmon

Research output: Contribution to journalArticle

32 Citations (Scopus)

Abstract

Previous studies have demonstrated a Factor IX and IXa binding site on the endothelial cell surface for which both the zymogen and enzyme compete with equal affinity. In this report, we demonstrate that the affinity of Factor IXa, but not Factor IX, for the cell surface is increased in the presence of both Factors VIII and X. When Factor Xa formation was studied in the presence of saturating concentrations of Factors VIII and X, the half-maximal rate was observed at a Factor IXa concentration of 151 ± 12 pM. Active site-blocked Factor IXa, 5-dimethylaminonaphthalene-1-sulfonyl-Glu-Gly-Arg-Factor IXa, was a more effective inhibitor of Factor X activation (K(i) = 124 pM) than was Factor IX (K(i) = 3.0 nM). Radioligand binding studies carred out in the presence of Factors VIII and X confirmed the presence of a selective endothelial cell Factor IXa binding site with K(d) = 127 ± 27 pM. In contrast, when Factor IXa binding was studied in the absence of other coagulation factors, or in the presence of Factor VIII (thrombin-activated or unactivated) alone, this new high affinity site was not observed. Competitive binding studies indicated that Factor IXa was 12 times more effective as an inhibitor of Factor IX-endothelial cell binding in the presence of Factors VIII and X. Consistent with the increased affinity of Factor IXa binding in the presence of factors VIII and X, cell-associated Factor IXa coagulant activity decayed 7 times more slowly in the presence of these coagulation factors. These results demonstrate selective Factor-IXa-endothelial cell binding in the presence of Factors VIII and X, suggesting this interaction could be a physiologic occurrence.

Original languageEnglish (US)
Pages (from-to)6717-6722
Number of pages6
JournalJournal of Biological Chemistry
Volume260
Issue number11
StatePublished - 1985
Externally publishedYes

Fingerprint

Factor IXa
Endothelial cells
Factor X
Endothelial Cells
Factor VIII
Factor IX
Blood Coagulation Factors
Factor VIIIa
Binding Sites
Coagulants
Enzyme Precursors
Factor Xa
Competitive Binding
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Stern, D., Nawroth, P. P., Kisiel, W., Vehar, G., & Esmon, C. T. (1985). The binding of factor IXa to cultured bovine aortic endothelial cells. Journal of Biological Chemistry, 260(11), 6717-6722.

The binding of factor IXa to cultured bovine aortic endothelial cells. / Stern, David; Nawroth, P. P.; Kisiel, W.; Vehar, G.; Esmon, C. T.

In: Journal of Biological Chemistry, Vol. 260, No. 11, 1985, p. 6717-6722.

Research output: Contribution to journalArticle

Stern, D, Nawroth, PP, Kisiel, W, Vehar, G & Esmon, CT 1985, 'The binding of factor IXa to cultured bovine aortic endothelial cells', Journal of Biological Chemistry, vol. 260, no. 11, pp. 6717-6722.
Stern, David ; Nawroth, P. P. ; Kisiel, W. ; Vehar, G. ; Esmon, C. T. / The binding of factor IXa to cultured bovine aortic endothelial cells. In: Journal of Biological Chemistry. 1985 ; Vol. 260, No. 11. pp. 6717-6722.
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