A prothrombinase complex of mouse peritoneal macrophages.
Addition of prothrombin to mouse peritoneal macrophages in vitro resulted in the formation of a thrombin-like enzyme, as demonstrated by use of the luminogenic peptide substrate S-2621. The prothrombinase activity was sedimented by high-speed centrifugation following homogenization of the cells and was abolished by treatment of the cells with the nonionic detergent Triton X-100 at 0.02% concentration. Moreover, the activity was drastically reduced by maintaining cultures in the presence of warfarin and, presumably due to competitive substrate inhibition, by adding S-2222, a chromogenic peptide substrate for Factor Xa. These findings suggest that prothrombin cleavage is catalyzed by Factor Xa at the macrophage surface. The generated thrombin was inhibited by antithrombin, and this reaction was accelerated by heparin with high affinity for antithrombin but not by the corresponding oligosaccharides composed of 8-14 monosaccharide units. Such oligosaccharides which are capable of accelerating the inactivation of Factor Xa by antithrombin, inhibited thrombin formation from prothrombin in the macrophage cultures, presumably by promoting inactivation by antithrombin of Factor Xa in a prothrombinase complex. Activation of the macrophage coagulation system, as proposed to occur in certain inflammatory conditions, thus may be modulated at various levels by heparin, or heparin oligosaccharides, released from mast cells.
Department of Veterinary Medical Chemistry, Biomedical Center, Uppsala, Sweden.
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Lindahl
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Pejler
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Bøgwald
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Seljelid
D000818:Animals; D001777:Blood Coagulation; D002462:Cell Membrane / Q000201:enzymology; D005165:Factor V / Q000032:analysis*; D005170:Factor X / Q000032:analysis*; D015951:Factor Xa; D008264:Macrophages / Q000201:enzymology* / Q000648:ultrastructure; D051379:Mice; D010529:Peritoneal Cavity / Q000166:cytology; D011516:Prothrombin / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism; D014859:Warfarin / Q000494:pharmacology
C022475:prothrombinase complex; D014859:Warfarin; D005165:Factor V; D011516:Prothrombin; D005170:Factor X; D012697:Serine Endopeptidases; D013917:Thrombin; D015951:Factor Xa