Bb. Sorensen et Lvm. Rao, INTERACTION OF ACTIVATED FACTOR-VII AND ACTIVE SITE-INHIBITED ACTIVATED FACTOR-VII WITH TISSUE FACTOR, Blood coagulation & fibrinolysis, 9, 1998, pp. 67-71
The coagulation cascade is initiated by binding of plasma activated or
non-activated factor VII [FVII(a)] to cell surface tissue factor (TF)
. TF-induced coagulation plays a primary role not only in haemostasis
but also in the pathogenesis of various thrombotic disorders. Recent s
tudies with animal model systems showed that the administration of act
ive site-inhibited FVIIa (FVIIai) blocked TF-FVIIa-induced fibrin and
thrombus formation. These data suggest that FVIIai competes with plasm
a FVII(a) for a limited number of TF sites expressed on cells either c
onstitutively or induced after the perturbation. To obtain insights in
to the mechanism(s) by which FVIIai is effective in inhibiting TF-FVII
a induced coagulation in vivo, we compared the interaction of FVIIai a
nd FVIIa with TF using a variety of competition assays and direct bind
ing assays. The TF-FVIIa amidolytic activity competition assay showed
that FVIIai bound with a threefold higher affinity than that of FVIIa
to TF relipidated in phosphatidylcholine (PC) vesicles, whereas no sig
nificant differences were found between FVIIa and FVIIai binding to TF
if it had been relipidated in mixed phospholipid vesicles containing
PC and phosphatidylserine (PS). When FVIIa and FVIIai binding to TF wa
s analysed in a FXa generation assay, we found that FVIIai bound to TF
in PCPS vesicles with two- to fivefold higher affinity than that of F
VIIa, whereas the affinity of FVIIai for TF in PC vesicles was seven-
to 10-fold higher than that of FVIIa. Direct binding analysis to TF, i
mmobilized on a sensor chip or on a cell surface, showed a faster asso
ciation and a slower dissociation of FVIIai to TF compared with that o
f FVIIa. Equilibrium binding to cell surface TF showed that the affini
ty of FVIIai was fivefold higher than that of FVIIa to non-functional
TF, whereas both FVIIa and FVIIai bound functional TF with the same hi
gh affinity. The enhanced affinity of FVII to TF, particularly to non-
functional TF, would make FVIIai a valuable reagent to block TF-induce
d coagulation before it is triggered by cell injury or a pathological
stimuli. Blood Coag Fibrinol 9 (suppl 1):S67-S71 (C) 1998 Lippincott-R
aven Publishers