Role of the Gla and first epidermal growth factor-like domains of factor X in the prothrombinase and tissue factor-factor VIIa complexes.
Factor X (FX) has high structure homology with other proteins of blood coagulation such as factor IX (FIX) and factor VII (FVII). These proteins present at their amino-terminal extremity a gamma-carboxyglutamic acid containing domain (Gla domain), followed by two epidermal growth factor-like (EGF1 and EGF2) domains, an activation peptide, and a serine protease domain. After vascular damage, the tissue factor-FVIIa (TF-FVIIa) complex activates both FX and FIX. FXa interacts stoichiometrically with tissue pathway inhibitor (TFPI), regulating TF-FVIIa activity by forming the TF-FVIIa-TFPI-FXa quaternary complex. Conversely, FXa boosts coagulation by its association with its cofactor, factor Va (FVa). To investigate the contribution of the Gla and EGF1 domains of FX in these complexes, FX chimeras were produced in which FIX Gla and EGF1 domains substituted the corresponding domains of FX. The affinity of the two chimeras, FX/FIX(Gla) and FX/FIX(EGF1), for the TF-FVIIa complex was markedly reduced compared with that of wild-type-FX (wt-FX) independently of the presence of phospholipids. Furthermore, the association rate constants of preformed FX/FIX(Gla)-TFPI and FX/FIX(EGF1)-TFPI complexes with TF-FVIIa were, respectively, 10- and 5-fold slower than that of wt-FXa-TFPI complex. Finally, the apparent affinity of FVa was 2-fold higher for the chimeras than for wt-FX in the presence of phospholipids and equal in their absence. These data demonstrate that FX Gla and EGF1 domains contain residues, which interact with TF-FVIIa exosites contributing to the formation of the TF-FVIIa-FX and TF-FVIIa-TFPI-FXa complexes. On the opposite, FXa Gla and EGF1 domains are not directly involved in FVa binding.
Thiec
Fabrice
F
INSERM U143, Hôpital Bicêtre, 94276 Le Kremlin-Bicêtre Cedex, France.
Cherel
Ghislaine
G
Christophe
Olivier D
OD
D015055:1-Carboxyglutamic Acid / Q000378:metabolism*; D000818:Animals; D004815:Epidermal Growth Factor / Q000378:metabolism*; D015942:Factor VIIa / Q000378:metabolism*; D005170:Factor X / Q000737:chemistry* / Q000502:physiology; D006801:Humans; D051379:Mice; D011516:Prothrombin / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D013329:Structure-Activity Relationship; D013925:Thromboplastin / Q000378:metabolism*
D011994:Recombinant Proteins; D015055:1-Carboxyglutamic Acid; D004815:Epidermal Growth Factor; D011516:Prothrombin; D005170:Factor X; D013925:Thromboplastin; D015942:Factor VIIa