scholarly journals Enhanced Rate of Cleavage at Arg-306 and Arg-506 in Coagulation Factor Va by Gla Domain-mutated Human-activated Protein C

2004 ◽  
Vol 279 (46) ◽  
pp. 47528-47535 ◽  
Author(s):  
Yong-Hui Sun ◽  
Sinh Tran ◽  
Eva A. Norstrøm ◽  
Björn Dahlbäck
Keyword(s):  
Protein C ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Factor Va ◽  
Gla Domain

scholarly journals Prothrombin amino terminal region helps protect coagulation factor Va from proteolytic inactivation by activated protein C

2009 ◽  
Vol 101 (01) ◽  
pp. 55-61 ◽  
Author(s):  
Subramanian Yegneswaran ◽  
Phuong Nguyen ◽  
John Griffin ◽  
Andrew Gale
Keyword(s):  
Protein Interactions ◽  
Protein C ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Critical Micellar Concentration ◽  
General Concept ◽  
Phospholipid Vesicle ◽  
Amino Terminal ◽  
Factor Va ◽  
Gla Domain

SummaryThe hypothesis that prothrombin (FII) protects coagulation factor Va (FVa) from proteolytic inactivation by activated protein C (APC) was tested using purified proteins. FII dose-dependently protected FVa from APC proteolysis under conditions where competition of proteins for binding to negatively-charged phospholipid surface was not relevant (i.e. either at high phospholipid vesicle concentrations or using soluble dicaproylphosphatidylserine at levels below its critical micellar concentration). Cleavages in FVa at both Arg506 and Arg306 by APC were inhibited by FII. FII did not alter the amidolytic activity of APC towards chromogenic oligopeptide substrates or inhibit FVIIIa inactivation by APC, implying that the FII-mediated protection of FVa from APC proteolysis was due to the ability of FII to inhibit protein-protein interactions between FVa and APC. FII also protected FVa from inactivation by Gla-domainless APC, ruling out a role for the APC Gla domain for these observations. To identify domains of FII responsible for the observed phenomenon, various forms or fragments of FII were employed. Biotin-PheProArg-CMK-inhibited meizothrombin and fII-fragment 1•2 protected FVa from proteolysis by APC. In contrast, no significant protection of FVa from APC cleavage was observed for Gladomainless-FII, prethrombin-1, prethrombin-2, FII fragment 1 or active site inhibited-thrombin (DEGR-thrombin). Overall, these data demonstrate that the Gla domain of FII linked to kringle 1 and 2 is necessary for the ability of FII to protect FVa from APC cleavage and support the general concept that assembly of the FII activation complex (FXa•FVa•FII•lipid surface) protects FVa from APC inactivation so that the procoagulant, thrombin generating pathway can act unhindered by APC. Only following FII activation and dissociation of the FII Gla domain fragments from the FII-ase complex, can APC inactivate FVa and down-regulate thrombin generation.

C4b-Binding Protein Protects Coagulation Factor Va from Inactivation by Activated Protein C†

Biochemistry ◽  
2000 ◽  
Vol 39 (47) ◽  
pp. 14543-14548 ◽  
Author(s):  
Robbert H. L. van de Poel ◽  
Joost C. M. Meijers ◽  
Jan Rosing ◽  
Guido Tans ◽  
Bonno N. Bouma
Keyword(s):  
Protein C ◽  
Binding Protein ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Factor Va

scholarly journals Interdomain engineered disulfide bond permitting elucidation of mechanisms of inactivation of coagulation factor Va by activated protein C

2009 ◽  
Vol 11 (9) ◽  
pp. 2091-2101 ◽  
Author(s):  
Andrew J. Gale ◽  
Xiao Xu ◽  
Jean-Luc Pellequer ◽  
Elizabeth D. Getzoff ◽  
John H. Griffin
Keyword(s):  
Disulfide Bond ◽  
Protein C ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Factor Va

scholarly journals Blood coagulation factor Va abnormality associated with resistance to activated protein C in venous thrombophilia

Blood ◽  
1994 ◽  
Vol 83 (11) ◽  
pp. 3120-3125 ◽  
Author(s):  
X Sun ◽  
B Evatt ◽  
JH Griffin
Keyword(s):  
Factor Viii ◽  
Protein C ◽  
Anticoagulant Activity ◽  
Laboratory Finding ◽  
Activated Protein C ◽  
Coagulation Factor ◽  
Factor V ◽  
Apc Resistance ◽  
Factor Va ◽  
Common Laboratory

Abstract A coagulation test abnormality, termed activated protein C (APC) resistance, involving poor anticoagulant response to APC is currently the most common laboratory finding among venous thrombophilic patients. Because the anticoagulant activity of APC involves inactivation of factors Va and VIIIa, studies were made to assess the presence of abnormal factors V or VIII. Diluted aliquots of plasma from two unrelated patients with APC resistance and thrombosis were added to either factor VIII-deficient or factor V-deficient plasma and APC resistance assays were performed. The results suggested that patients' factor V but not factor VIII rendered the substrate plasma APC resistant. When factor V that had been partially purified from normal or APC resistant patients' plasmas using immunoaffinity chromatography was added to factor V-deficient plasma, APC resistance assays showed that patients' factor V or factor Va, but not normal factor V, rendered the substrate plasma resistant to APC. Studies of the inactivation of each partially purified thrombin-activated factor Va by APC suggested that half of the patients' factor Va was resistant to APC. These results support the hypothesis that the APC resistance of some venous thrombophilic plasmas is caused by abnormal factor Va.

Activated protein C cofactor function of protein S: a novel role for a γ-carboxyglutamic acid residue

Blood ◽  
2011 ◽  
Vol 117 (24) ◽  
pp. 6685-6693 ◽  
Author(s):  
Josefin Ahnström ◽  
Helena M. Andersson ◽  
Kevin Canis ◽  
Eva Norstrøm ◽  
Yao Yu ◽  
...  
Keyword(s):  
Protein C ◽  
Single Point ◽  
Activated Protein C ◽  
Protein S ◽  
Phospholipid Binding ◽  
Interaction Sites ◽  
Factor Va ◽  
Factor Viiia ◽  
Gla Domain ◽  
Anticoagulant Function

Abstract Protein S has an important anticoagulant function by acting as a cofactor for activated protein C (APC). We recently reported that the EGF1 domain residue Asp95 is critical for APC cofactor function. In the present study, we examined whether additional interaction sites within the Gla domain of protein S might contribute to its APC cofactor function. We examined 4 residues, composing the previously reported “Face1” (N33S/P35T/E36A/Y39V) variant, as single point substitutions. Of these protein S variants, protein S E36A was found to be almost completely inactive using calibrated automated thrombography. In factor Va inactivation assays, protein S E36A had 89% reduced cofactor activity compared with wild-type protein S and was almost completely inactive in factor VIIIa inactivation; phospholipid binding was, however, normal. Glu36 lies outside the ω-loop that mediates Ca2+-dependent phospholipid binding. Using mass spectrometry, it was nevertheless confirmed that Glu36 is γ-carboxylated. Our finding that Gla36 is important for APC cofactor function, but not for phospholipid binding, defines a novel function (other than Ca2+ coordination/phospholipid binding) for a Gla residue in vitamin K–dependent proteins. It also suggests that residues within the Gla and EGF1 domains of protein S act cooperatively for its APC cofactor function.

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