Factor V is a substrate for the transamidase factor XIIIa.
Coagulation Factor V (Mr = 330,000), upon cleavage by thrombin, produces Factor Va, which is composed of two subunits with Mr values of 94,000 and 74,000, along with two activation fragments possessing no known function. Studies were undertaken to assess the ability of the transamidase Factor XIIIa to covalently incorporate the lysine analogs [3H]putrescine and dansylcadaverine into the thrombin-cleaved (activated) and unactivated forms of human and bovine Factor V. The incorporation of either probe into thrombin-activated Factor V proceeded at an initial rate approximately twice that for unactivated Factor V. The extent of the incorporation of [3H]putrescine or dansylcadaverine into activated or unactivated human Factor V was identical; 4 mol of either probe per mol of Factor V. In the case of bovine Factor V, however, while 4 mol of probe were bound per mol of the unactivated pro-cofactor, 5 mol of either lysine analog were covalently linked to 1 mol of thrombin-cleaved Factor V. Polyacrylamide gel fluorography, immunoaffinity chromatography, and immunoprecipitation identified the largest activation fragment of human Factor V (Mr = 150,000) and bovine Factor V (Mr = 120,000) to contain the sites of incorporation of the covalently bound probes. High molecular weight, apparently covalent polymers of Factor V were produced by the action of Factor XIIIa on activated and unactivated human or bovine Factor V. The absence of either probe in the reaction mixtures did not appear to allow an enhancement of protein polymerization.
Francis
R T
RT
McDonagh
J
J
Mann
K G
KG
D000818:Animals; D002103:Cadaverine / Q000031:analogs & derivatives / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D004789:Enzyme Activation; D005165:Factor V / Q000378:metabolism*; D005176:Factor XIII / Q000378:metabolism*; D005470:Fluorometry; D007163:Immunosorbent Techniques; D008239:Lysine / Q000031:analogs & derivatives; D008970:Molecular Weight; D011700:Putrescine / Q000378:metabolism; D013917:Thrombin / Q000378:metabolism; D011503:Transglutaminases
D005165:Factor V; D005176:Factor XIII; D011503:Transglutaminases; D013917:Thrombin; C008542:monodansylcadaverine; D008239:Lysine; D002103:Cadaverine; D011700:Putrescine