The Factor IX γ-Carboxyglutamic Acid (Gla) Domain Is Involved in Interactions between Factor IX and Factor XIa
During hemostasis, factor IX is activated to factor IXaβ by factor VIIa and factor XIa. The glutamic acid-rich γ-carboxyglutamic acid (Gla) domain of factor IX is involved in phospholipid binding and is required for activation by factor VIIa. In contrast, activation by factor XIa is not phospholip...
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| Veröffentlicht in: | The Journal of biological chemistry 2003-03, Vol.278 (10), p.7981 |
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| creator | Aysar Aktimur Melanie A. Gabriel David Gailani John R. Toomey |
| description | During hemostasis, factor IX is activated to factor IXaβ by factor VIIa and factor XIa. The glutamic acid-rich γ-carboxyglutamic
acid (Gla) domain of factor IX is involved in phospholipid binding and is required for activation by factor VIIa. In contrast,
activation by factor XIa is not phospholipid-dependent, raising questions about the importance of the Gla for this reaction.
We examined binding of factors IX and IXaβ to factor XIa by surface plasmon resonance. Plasma factors IX and IXaβ bind to
factor XIa with K d values of 120 ± 11 n m and 110 ± 8 n m , respectively. Recombinant factor IX bound to factor XIa with a K d of 107 n m , whereas factor IX with a factor VII Gla domain (rFIX/VII-Gla) and factor IX expressed in the presence of warfarin (rFIX-desγ)
did not bind. An anti-factor IX Gla monoclonal antibody was a potent inhibitor of factor IX binding to factor XIa ( K i 34 n m ) and activation by factor XIa ( K i 33 n m ). In activated partial thromboplastin time clotting assays, the specific activities of plasma and recombinant factor IX were
comparable (200 and 150 units/mg), whereas rFIX/VII-Gla activity was low ( |
| doi_str_mv | 10.1074/jbc.M212748200 |
| format | Article |
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acid (Gla) domain of factor IX is involved in phospholipid binding and is required for activation by factor VIIa. In contrast,
activation by factor XIa is not phospholipid-dependent, raising questions about the importance of the Gla for this reaction.
We examined binding of factors IX and IXaβ to factor XIa by surface plasmon resonance. Plasma factors IX and IXaβ bind to
factor XIa with K d values of 120 ± 11 n m and 110 ± 8 n m , respectively. Recombinant factor IX bound to factor XIa with a K d of 107 n m , whereas factor IX with a factor VII Gla domain (rFIX/VII-Gla) and factor IX expressed in the presence of warfarin (rFIX-desγ)
did not bind. An anti-factor IX Gla monoclonal antibody was a potent inhibitor of factor IX binding to factor XIa ( K i 34 n m ) and activation by factor XIa ( K i 33 n m ). In activated partial thromboplastin time clotting assays, the specific activities of plasma and recombinant factor IX were
comparable (200 and 150 units/mg), whereas rFIX/VII-Gla activity was low (<2 units/mg). In contrast, recombinant factor IXaβ
and activated rFIX/VIIa-Gla had similar activities (80 and 60% of plasma factor IXaβ), indicating that both proteases activate
factor X and that the poor activity of zymogen rFIX/VII-Gla was caused by a specific defect in activation by factor XIa. The
data demonstrate that factor XIa binds with comparable affinity to factors IX and IXaβ and that the interactions are dependent
on the factor IX Gla domain.</description><identifier>ISSN: 0021-9258</identifier><identifier>EISSN: 1083-351X</identifier><identifier>DOI: 10.1074/jbc.M212748200</identifier><identifier>PMID: 12496253</identifier><language>eng</language><publisher>American Society for Biochemistry and Molecular Biology</publisher><ispartof>The Journal of biological chemistry, 2003-03, Vol.278 (10), p.7981</ispartof><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,776,780,27901,27902</link.rule.ids></links><search><creatorcontrib>Aysar Aktimur</creatorcontrib><creatorcontrib>Melanie A. Gabriel</creatorcontrib><creatorcontrib>David Gailani</creatorcontrib><creatorcontrib>John R. Toomey</creatorcontrib><title>The Factor IX γ-Carboxyglutamic Acid (Gla) Domain Is Involved in Interactions between Factor IX and Factor XIa</title><title>The Journal of biological chemistry</title><description>During hemostasis, factor IX is activated to factor IXaβ by factor VIIa and factor XIa. The glutamic acid-rich γ-carboxyglutamic
acid (Gla) domain of factor IX is involved in phospholipid binding and is required for activation by factor VIIa. In contrast,
activation by factor XIa is not phospholipid-dependent, raising questions about the importance of the Gla for this reaction.
We examined binding of factors IX and IXaβ to factor XIa by surface plasmon resonance. Plasma factors IX and IXaβ bind to
factor XIa with K d values of 120 ± 11 n m and 110 ± 8 n m , respectively. Recombinant factor IX bound to factor XIa with a K d of 107 n m , whereas factor IX with a factor VII Gla domain (rFIX/VII-Gla) and factor IX expressed in the presence of warfarin (rFIX-desγ)
did not bind. An anti-factor IX Gla monoclonal antibody was a potent inhibitor of factor IX binding to factor XIa ( K i 34 n m ) and activation by factor XIa ( K i 33 n m ). In activated partial thromboplastin time clotting assays, the specific activities of plasma and recombinant factor IX were
comparable (200 and 150 units/mg), whereas rFIX/VII-Gla activity was low (<2 units/mg). In contrast, recombinant factor IXaβ
and activated rFIX/VIIa-Gla had similar activities (80 and 60% of plasma factor IXaβ), indicating that both proteases activate
factor X and that the poor activity of zymogen rFIX/VII-Gla was caused by a specific defect in activation by factor XIa. The
data demonstrate that factor XIa binds with comparable affinity to factors IX and IXaβ and that the interactions are dependent
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acid (Gla) domain of factor IX is involved in phospholipid binding and is required for activation by factor VIIa. In contrast,
activation by factor XIa is not phospholipid-dependent, raising questions about the importance of the Gla for this reaction.
We examined binding of factors IX and IXaβ to factor XIa by surface plasmon resonance. Plasma factors IX and IXaβ bind to
factor XIa with K d values of 120 ± 11 n m and 110 ± 8 n m , respectively. Recombinant factor IX bound to factor XIa with a K d of 107 n m , whereas factor IX with a factor VII Gla domain (rFIX/VII-Gla) and factor IX expressed in the presence of warfarin (rFIX-desγ)
did not bind. An anti-factor IX Gla monoclonal antibody was a potent inhibitor of factor IX binding to factor XIa ( K i 34 n m ) and activation by factor XIa ( K i 33 n m ). In activated partial thromboplastin time clotting assays, the specific activities of plasma and recombinant factor IX were
comparable (200 and 150 units/mg), whereas rFIX/VII-Gla activity was low (<2 units/mg). In contrast, recombinant factor IXaβ
and activated rFIX/VIIa-Gla had similar activities (80 and 60% of plasma factor IXaβ), indicating that both proteases activate
factor X and that the poor activity of zymogen rFIX/VII-Gla was caused by a specific defect in activation by factor XIa. The
data demonstrate that factor XIa binds with comparable affinity to factors IX and IXaβ and that the interactions are dependent
on the factor IX Gla domain.</abstract><pub>American Society for Biochemistry and Molecular Biology</pub><pmid>12496253</pmid><doi>10.1074/jbc.M212748200</doi></addata></record> |
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| source | EZB-FREE-00999 freely available EZB journals; Alma/SFX Local Collection |
| title | The Factor IX γ-Carboxyglutamic Acid (Gla) Domain Is Involved in Interactions between Factor IX and Factor XIa |
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