scholarly journals Identification of the Phospholipid Binding Site in the Vitamin K-dependent Blood Coagulation Protein Factor IX

1996 ◽  
Vol 271 (27) ◽  
pp. 16227-16236 ◽  
Author(s):  
Steven J. Freedman ◽  
Mark D. Blostein ◽  
James D. Baleja ◽  
Margaret Jacobs ◽  
Barbara C. Furie ◽  
...  
Keyword(s):  
Binding Site ◽  
Blood Coagulation ◽  
Vitamin K ◽  
Factor Ix ◽  
Phospholipid Binding ◽  
Protein Factor ◽  
Coagulation Protein

A MONOCLONAL ANTIBODY WHOSE BINDING IS INHIBITED BY DIVALENT CATIONS

1987 ◽  
Author(s):  
M de Serres ◽  
H M Reisner ◽  
D Monroe ◽  
H Roberts
Keyword(s):  
Monoclonal Antibody ◽  
Vitamin K ◽  
Divalent Cations ◽  
Factor Ix ◽  
Protein Preparation ◽  
Coagulant Activity ◽  
Maximum Inhibition ◽  
Standard Assay ◽  
Coagulation Protein ◽  
Direct Binding

Factor IX (FIX), a vitamin K dependent coagulation protein, is functionally deficient or absent in patients with hemophilia B. Binding of Ca++ by the gammacarboxyglutamic acid residues (Gla) of FIX is necessary for coagulant activity. Antibodies havebeendes-cribed which selectively bind to FIX in the presence of Ca++ and appear to interact with Ca-H- stabilized epitopes of FIX. One IgM, Kappa murine monoclonal antibody (Mab), 129-1, has been found to react preferentially with FIX in the absence of Ca-H- or with FIX having a reduction in gammacarboxylation. 129-1 was characterized by direct binding ELISA assays, the standard assay buffer (.02M Tris - .15M NaCL pH 7.5) was supplemented with Ca-H- or ED TA to final concentrations to 5mM and lOmM respectively. In the presence of Ca-H-, titers ranged from 10 to 100 as compared to 100 K to 500 K in the presence of EDTA. Control Mab's, FIX-30 and 2D521 showed no such effect. Maximum inhibition occurred ≥2.5 mM Ca-H- concentration. Mg++, Sr-H- and Mn-H- were tested with similar results. Chemically degammacarboxylated FIXa was compared to FIX and FIXa controls in the presence of Ca-H- or EDTA, to determine the importance of Gla residues. In the presence of EDTA, Mab 129-1 had essentially equal reactivity with all these proteins (titers 100 K). In the presence of Ca-H-, binding to FIX and FIXa was inhibited (500 and 1 K respectively). However, the binding to Gla modified FIXa in the presence of Ca-H- was only slightly reduced (10 K), suggesting the importance of Gla residues in the Ca-H- mediated inhibition. FIX was purified from concentrate, coumadinized plasma and culture supernatant (produced in the absence of vitamin K) from the cell line PMN45 using HPLC affinity chromatography with Mab 2D521. Mab 129-1 binding to FIX immunopurified from concentrate and a control biochemically purified FIX protein preparation was significantly higher in the presence of EDTA (titers 500 and 5 K) than in Ca-H- (titers 10 and 10). FIX purified from coumadinized plasma or PMN45 cells showed equal low reactivity with 129-1 in the presence or absence of Ca-H- (titer 10 in all cases). FIX-30 and 2D521 controls showed no such reduction in binding. Therefore, Gla residues may play a role in the binding of 129-1 to FIX.

scholarly journals The Sequence GluLys of Human Blood Coagulation Factor VIII Comprises a Binding Site for Activated Factor IX

1996 ◽  
Vol 271 (4) ◽  
pp. 1935-1940 ◽  
Author(s):  
Peter J. Lenting ◽  
Jan-Willem H. P. van de Loo ◽  
Marie-José S. H. Donath ◽  
Jan A. van Mourik ◽  
Koen Mertens
Keyword(s):  
Factor Viii ◽  
Binding Site ◽  
Blood Coagulation ◽  
Human Blood ◽  
Coagulation Factor ◽  
Factor Ix ◽  
Coagulation Factor Viii ◽  
Blood Coagulation Factor Viii ◽  
Human Blood Coagulation

Localization of the Factor IX Propeptide Binding Site on Recombinant Vitamin K Dependent Carboxylase Using Benzoylphenylalanine Photoaffinity Peptide Inactivators

Biochemistry ◽  
1995 ◽  
Vol 34 (2) ◽  
pp. 481-489 ◽  
Author(s):  
Masahiko Yamada ◽  
Athan Kuliopulos ◽  
Noele P. Nelson ◽  
David A. Roth ◽  
Bruce Furie ◽  
...  
Keyword(s):  
Binding Site ◽  
Vitamin K ◽  
Factor Ix

Current perspectives in antithrombotic therapy

2001 ◽  
Vol 21 (03) ◽  
pp. 82-96 ◽  
Author(s):  
D. Hoppensteadt ◽  
O. Iqbal ◽  
R. L. Bick ◽  
J. Fareed
Keyword(s):  
Blood Coagulation ◽  
Antithrombotic Therapy ◽  
Thrombotic Event ◽  
The United States ◽  
Vascular Thrombosis ◽  
Common Cause ◽  
Coagulation Protein ◽  
Coronary Events ◽  
Platelet Defects

SummaryThrombotic disorders are the most common cause of death in the United States. About two million individuals die each year from an arterial or venous thrombosis or related disorders. About 80% to 90% of all cases of thrombosis can now be defined with respect to cause. Of these, over 50% occur in patients who harbor a congenital or acquired blood coagulation protein or platelet defect which caused the thrombotic event. It is obviously of major importance to define those individuals harboring such a defect as this allows: 1) appropriate antithrombotic therapy to decrease risks of recurrence; 2) determination of the length of time the patient must remain on therapy for secondary prevention; and 3) allow for testing of family members of those harboring a blood coagulation protein or platelet defect which is hereditary (about 50% of all coagulation and platelet defects mentioned above). Aside from mortality, significant additional morbidity occurs from both arterial or venous thrombotic events, including, but not limited to paralysis (non-fatal thrombotic stroke), cardiac disability (repeated coronary events), loss of vision (retinal vascular thrombosis), fetal waste syndrome (placental vascular thrombosis), stasis ulcers and other manifestations of post-phlebitic syndrome, etc.

Sign in / Sign up

Export Citation Format

Share Document