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Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites by Jialan Shi, and Gary E. Gilbert Blood Volume.

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Presentation on theme: "Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites by Jialan Shi, and Gary E. Gilbert Blood Volume."— Presentation transcript:

1 Lactadherin inhibits enzyme complexes of blood coagulation by competing for phospholipid-binding sites by Jialan Shi, and Gary E. Gilbert Blood Volume 101(7):2628-2636 April 1, 2003 ©2003 by American Society of Hematology

2 Competition for factor VIII– and factor V–binding sites by lactadherin or annexin V.(A) First, 4 nM fluorescein-labeled factor VIII was mixed with the indicated concentration of competitor, lactadherin ( ▴ ) or annexin V (●), in the presence of 1.5 mM Ca++. Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

3 Relationship of vesicle curvature and PS content to inhibition of the prothrombinase complex by lactadherin or annexin V.Sonicated vesicles ( ▴ and ▵ ), extruded vesicles (● and ○), and LMVs (▪ and ■) contained 4% PS (top row) or 15% PS (bottom row). Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

4 Relationship of vesicle curvature and PS content to inhibition of the factor Xase complex by lactadherin or annexin V.Sonicated vesicles ( ▴ and ▵ ), extruded vesicles (● and ○), and LMVs (▪ and ■) contained 4% PS (upper row) or 15% PS (lower row). Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

5 Inhibition of the factor VIIa–tissue factor complex by lactadherin or annexin V.Recombinant tissue factor was reconstituted into vesicles of PS-PE-PC composition of 4:20:76 by detergent dialysis, with a tissue factor-PL ratio of 1:7500. Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

6 Competition for factor IXa–binding sites by lactadherin.The active site histidine of factor IXa was derivatized with fluorescein-Glu-Gly-Arg chloromethyl ketone as described in “Materials and methods.” First, 4 nM fluorescein-labeled factor IXa was mixed wi... Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

7 Relationship of inhibition by lactadherin to phospholipid concentration.The function of factor Xase complex (A) and prothrombinase complex (B) on sonicated vesicles was evaluated at 0.1 (●), 0.5 (■), and 2.5 ( ▴ ) μM phospholipid concentrations in the presenc... Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology

8 Inhibition of whole blood prothrombin time by lactadherin or annexin V.Fresh whole blood was anticoagulated with 10 mM citrate and 25 μg/mL corn trypsin inhibitor. Jialan Shi, and Gary E. Gilbert Blood 2003;101:2628-2636 ©2003 by American Society of Hematology


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