Covalent crosslinking of human coagulation factor V by activated factor XIII from guinea pig megakaryocytes and human plasma.
Coagulation factor V (FV) has been shown to be synthesized in both the liver and megakaryocytes. We now present evidence that FV can be covalently crosslinked by an enzyme originating from megakaryocytes to form polymeric multimers of factor V. The guinea pig megakaryocyte enzyme appears to be factor XIIIa since the FV-crosslinking activity (1) had an absolute requirement for Ca++, (2) was completely inhibited by iodoacetamide, 5,5'-dithiobis- (2-nitrobenzoic acid), p-chloromercuribenzene sulfonic acid, and N-ethylmaleimide, all known alkylators of the thiol group at the active site of the factor XIIIa, (3) was blocked by known pseudoamine donor substrates of factor XIIIa including dansylcadaverine and putrescine, and (4) could be directly demonstrated in the guinea pig megakaryocyte lysate by a specific activity staining procedure. No tranglutaminase was detected in guinea pig megakaryocytes in contrast to red cells and liver. A similar pattern of covalent crosslinking of human FV by purified activated human plasma factor XIII was also demonstrated. Analysis of the crosslinked products of FV formed by the guinea pig enzyme by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) indicates the formation of intermediate as well as higher molecular weight polymers, suggesting that the crosslinking is a stepwise polymerization process.
Huh
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MM
Department of Medicine, Temple University School of Medicine, Philadelphia 19140.
Schick
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Schick
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Colman
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D000588:Amines / Q000494:pharmacology; D000818:Animals; D002118:Calcium / Q000378:metabolism; D003432:Cross-Linking Reagents; D005165:Factor V / Q000378:metabolism*; D005176:Factor XIII / Q000378:metabolism*; D006168:Guinea Pigs; D006801:Humans; D066298:In Vitro Techniques; D008533:Megakaryocytes / Q000378:metabolism*; D008970:Molecular Weight; D011108:Polymers; D013439:Sulfhydryl Reagents / Q000494:pharmacology
D000588:Amines; D003432:Cross-Linking Reagents; D011108:Polymers; D013439:Sulfhydryl Reagents; D005165:Factor V; D005176:Factor XIII; D002118:Calcium