Structural determinants of the factor IX molecule mediating interaction with the endothelial cell binding site are distinct from those involved in phospholipid binding

J. Ryan, B. Wolitzky, E. Heimer, T. Lambrose, A. Felix, J. P. Tam, L. H. Huang, P. Nawroth, G. Wilner, W. Kisiel, G. L. Nelsestuen, David Stern

Research output: Contribution to journalArticle

16 Citations (Scopus)

Abstract

Previous studies have indicated that Factor IX/IXa interacts in a specific and high affinity manner with a binding site on the endothelial cell surface. In this study, the contributions of the γ-carboxyglutamic acid-containing (GLA) and growth factor domains to the finding of Factor IX to the endothelium were assessed. While GLA-containing peptides from Factors IX, X, and prothrombin were inhibitors of 125I-Factor IX-endothelial cell binding, the GLA peptide from Factor IX was about 250-800-fold more effective than those from prothrombin and Factor X, respectively. In contrast to its relative efficacy as an inhibitor of Factor IX-cell surface interaction, the Factor IX-GLA peptide neither bound to lipid vesicles nor inhibited Factor IX-lipid interaction. A synthetic peptide comprising the entire first epidermal growth factor (EGF) exon was also an inhibitor of 125I-Factor IX-endothelial cell binding, although it did not interact with lipid vesicles. Experiments with synthetic peptides comprising each of the three loops of the first EGF domain or the entire first EGF region with specific substitutions indicated the importance of determinants in both the first and probably third loops for Factor IX-endothelial interaction. In contrast, the second loop of the first EGF domain and the first loop of the second EGF exon are probably not involved in Factor IX-endothelial interaction based on their inability to block 125I-Factor IX binding to cells. These results indicate that determinants in both the GLA and the first EGF domain contribute to the specific binding of Factor IX to the endothelial cell surface and that structural requirements for Factor IX-cell surface interaction are distinct from those for Factor IX binding to lipids.

Original languageEnglish (US)
Pages (from-to)20283-20287
Number of pages5
JournalJournal of Biological Chemistry
Volume264
Issue number34
StatePublished - 1989
Externally publishedYes

Fingerprint

Factor IX
Endothelial cells
Phospholipids
Endothelial Cells
Binding Sites
Molecules
Epidermal Growth Factor
Lipids
Prothrombin
Cell Communication
Peptides
Exons
Factor IXa
Factor X
Endothelium
Intercellular Signaling Peptides and Proteins
Substitution reactions

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Structural determinants of the factor IX molecule mediating interaction with the endothelial cell binding site are distinct from those involved in phospholipid binding. / Ryan, J.; Wolitzky, B.; Heimer, E.; Lambrose, T.; Felix, A.; Tam, J. P.; Huang, L. H.; Nawroth, P.; Wilner, G.; Kisiel, W.; Nelsestuen, G. L.; Stern, David.

In: Journal of Biological Chemistry, Vol. 264, No. 34, 1989, p. 20283-20287.

Research output: Contribution to journalArticle

Ryan, J, Wolitzky, B, Heimer, E, Lambrose, T, Felix, A, Tam, JP, Huang, LH, Nawroth, P, Wilner, G, Kisiel, W, Nelsestuen, GL & Stern, D 1989, 'Structural determinants of the factor IX molecule mediating interaction with the endothelial cell binding site are distinct from those involved in phospholipid binding', Journal of Biological Chemistry, vol. 264, no. 34, pp. 20283-20287.
Ryan, J. ; Wolitzky, B. ; Heimer, E. ; Lambrose, T. ; Felix, A. ; Tam, J. P. ; Huang, L. H. ; Nawroth, P. ; Wilner, G. ; Kisiel, W. ; Nelsestuen, G. L. ; Stern, David. / Structural determinants of the factor IX molecule mediating interaction with the endothelial cell binding site are distinct from those involved in phospholipid binding. In: Journal of Biological Chemistry. 1989 ; Vol. 264, No. 34. pp. 20283-20287.
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abstract = "Previous studies have indicated that Factor IX/IXa interacts in a specific and high affinity manner with a binding site on the endothelial cell surface. In this study, the contributions of the γ-carboxyglutamic acid-containing (GLA) and growth factor domains to the finding of Factor IX to the endothelium were assessed. While GLA-containing peptides from Factors IX, X, and prothrombin were inhibitors of 125I-Factor IX-endothelial cell binding, the GLA peptide from Factor IX was about 250-800-fold more effective than those from prothrombin and Factor X, respectively. In contrast to its relative efficacy as an inhibitor of Factor IX-cell surface interaction, the Factor IX-GLA peptide neither bound to lipid vesicles nor inhibited Factor IX-lipid interaction. A synthetic peptide comprising the entire first epidermal growth factor (EGF) exon was also an inhibitor of 125I-Factor IX-endothelial cell binding, although it did not interact with lipid vesicles. Experiments with synthetic peptides comprising each of the three loops of the first EGF domain or the entire first EGF region with specific substitutions indicated the importance of determinants in both the first and probably third loops for Factor IX-endothelial interaction. In contrast, the second loop of the first EGF domain and the first loop of the second EGF exon are probably not involved in Factor IX-endothelial interaction based on their inability to block 125I-Factor IX binding to cells. These results indicate that determinants in both the GLA and the first EGF domain contribute to the specific binding of Factor IX to the endothelial cell surface and that structural requirements for Factor IX-cell surface interaction are distinct from those for Factor IX binding to lipids.",
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AU - Wolitzky, B.

AU - Heimer, E.

AU - Lambrose, T.

AU - Felix, A.

AU - Tam, J. P.

AU - Huang, L. H.

AU - Nawroth, P.

AU - Wilner, G.

AU - Kisiel, W.

AU - Nelsestuen, G. L.

AU - Stern, David

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