Human annexin V binds to sulfatide: contribution to regulation of blood coagulation.
Annexin V is a calcium-dependent phospholipid-binding protein that exhibits anticoagulant activity on binding to phosphatidylserine exposed on the activated surfaces of endothelial cells and platelets, inhibiting activation of factor X and prothrombin in the blood coagulation cascade. Sulfatide (galactosylceramide I(3)-sulfate), one of the glycosphingolipids of the platelet cell membrane, is thought to be involved in blood coagulation systems via activation of factor XII. In this study, we examined whether or not annexin V binds to sulfatide and affects the coagulant activity of sulfatide. Solid phase assaying of annexin V revealed that it binds specifically to sulfatide, i.e. not to galactosylceramide or gangliosides, in the presence of calcium ions. Affinity analysis by means of surface plasmon resonance showed that the K(D) of the interaction between annexin V and sulfatide is 1.2 micro M. Kinetic turbidometric assaying of plasma coagulation initiated by CaCl(2) revealed that the coagulation rate in the presence of sulfatide or phosphatidylserine was decreased by annexin V. These results suggest that annexin V regulates coagulability in the blood stream by binding not only to phosphatidylserine but also to sulfatide.
Ida
Michiru
M
The Graduate School of Humanities and Sciences, Ochanomizu University, 2-1-1 Otsuka, Bunkyo-ku, Tokyo 112-8610.
Satoh
Ayano
A
Matsumoto
Isamu
I
Kojima-Aikawa
Kyoko
K
D017304:Annexin A5 / Q000737:chemistry* / Q000378:metabolism / Q000494:pharmacology; D001777:Blood Coagulation; D002118:Calcium / Q000378:metabolism; D002122:Calcium Chloride / Q000494:pharmacology; D002855:Chromatography, Thin Layer; D004305:Dose-Response Relationship, Drug; D005170:Factor X / Q000037:antagonists & inhibitors; D005699:Galactosylceramides / Q000378:metabolism; D005732:Gangliosides / Q000378:metabolism; D005982:Glutathione Transferase / Q000378:metabolism; D006017:Glycolipids / Q000737:chemistry; D006801:Humans; D007477:Ions; D007700:Kinetics; D037102:Lectins / Q000737:chemistry; D011485:Protein Binding; D011516:Prothrombin / Q000037:antagonists & inhibitors; D011994:Recombinant Proteins / Q000737:chemistry; D013433:Sulfoglycosphingolipids / Q000737:chemistry*; D020349:Surface Plasmon Resonance; D013997:Time Factors
D017304:Annexin A5; D005699:Galactosylceramides; D005732:Gangliosides; D006017:Glycolipids; D007477:Ions; D037102:Lectins; D011994:Recombinant Proteins; D013433:Sulfoglycosphingolipids; D011516:Prothrombin; D005170:Factor X; D005982:Glutathione Transferase; D002122:Calcium Chloride; D002118:Calcium