Binding of annexin V to a human ovarian carcinoma cell line (OC-2008). Contrasting effects on cell surface factor VIIa/tissue factor activity and prothrombinase activity.
Proteins of the annexin/lipocortin family bind tightly to anionic phospholipids and platelets and act as in vitro anticoagulants. Annexins may be useful as tools to study the availability of anionic phospholipids on cell surfaces and their role in the regulation of blood coagulation. In the present study, we investigated the binding of annexin V (placental anticoagulant protein I) to a human ovarian carcinoma cell line, OC-2008, that constitutively expresses surface membrane tissue factor activity. Binding of annexin V to cell monolayers was calcium-dependent, specific, saturable and reversible; Scatchard analysis indicated a single class of binding sites with an apparent Kd of 9.4 +/- 3.1 nM and 5.2 +/- 1 x 10(6) sites per cell. Binding was completely inhibited by phospholipid vesicles containing phosphatidylserine, but was not inhibited by vesicles containing phosphatidylcholine. Annexin V inhibited the cell surface-dependent activity of prothrombinase complex, but did not inhibit the activity of the factor VIIa/tissue factor complex. In conclusion, these results suggest that anionic phospholipid is present on the extracellular face of OC-2008 cells; this anionic phospholipid is functionally important for the activity of the prothrombinase complex, but the importance of anionic phospholipid for the cell surface factor VIIa/tissue factor functional activity is unclear.
Rao
L V
LV
Department of Medicine, University of California, San Diego 92093.
Tait
J F
JF
Hoang
A D
AD
D017304:Annexin A5 / Q000378:metabolism* / Q000494:pharmacology; D001665:Binding Sites; D001777:Blood Coagulation; D015942:Factor VIIa / Q000378:metabolism; D005260:Female; D006801:Humans; D009097:Multienzyme Complexes; D010051:Ovarian Neoplasms / Q000378:metabolism*; D010718:Phosphatidylserines / Q000494:pharmacology; D013925:Thromboplastin / Q000037:antagonists & inhibitors* / Q000737:chemistry; D014407:Tumor Cells, Cultured
D017304:Annexin A5; D009097:Multienzyme Complexes; D010718:Phosphatidylserines; D013925:Thromboplastin; D015942:Factor VIIa