Carbohydrate residues modulate the activation of coagulation factor X.
Factor X is a plasma protein involved in both the intrinsic and extrinsic pathways of blood coagulation. Post-translational modifications of the protein involve gamma-carboxylation of specific glutamic acid residues, beta-hydroxylation of one aspartic acid residue, and N- and O-linked glycosylation. Even though it is known that gamma-carboxylation is instrumental in regulating biological activity, the role of glycosylation in the function and properties of factor X has not been previously investigated. We utilized lectin binding and glycosidase treatment to investigate the functional role of carbohydrates on the activation peptide of factor X. Sambucus nigra agglutinin, a lectin that binds to sialic acid terminally linked alpha(2-6) to galactose or N-acetyl-galactosamine inhibits activation of human factor X in a dose-dependent manner. Inhibition of activation was observed for both intrinsic (factor IXa/VIIIa) and extrinsic (factor VIIa/tissue factor) pathway complexes. In accordance with this, selective removal of sialic acid residues on the activation peptide of factor X by neuraminidase also results in a drastic reduction of activation of the zymogen by these complexes. Corresponding reduction of activity in classical clotting assays (activated partial thromboplastin time and prothrombin time) also agrees with this observation. These results suggest a possible role of N-linked carbohydrates in the activation of factor X.
Sinha
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COR Therapeutics Inc., South San Francisco, California 94080.
Wolf
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DL
D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology*; D002417:Cattle; D004591:Electrophoresis, Polyacrylamide Gel; D015949:Factor IXa / Q000378:metabolism; D015942:Factor VIIa / Q000378:metabolism; D005170:Factor X / Q000037:antagonists & inhibitors / Q000302:isolation & purification / Q000378:metabolism*; D015951:Factor Xa / Q000378:metabolism*; D006026:Glycoside Hydrolases; D006801:Humans; D007700:Kinetics; D037102:Lectins / Q000494:pharmacology*; D008970:Molecular Weight; D013925:Thromboplastin / Q000378:metabolism
D002241:Carbohydrates; D037102:Lectins; D005170:Factor X; D013925:Thromboplastin; D006026:Glycoside Hydrolases; D015942:Factor VIIa; D015949:Factor IXa; D015951:Factor Xa