Catalytic life of activated factor XIII in thrombi. Implications for fibrinolytic resistance and thrombus aging.
BACKGROUND
Because the increased fibrinolytic resistance of older thrombi may be caused by the continuous cross-linking action of fibrin-bound activated factor XIII (FXIIIa), we examined the persistence of FXIIIa catalytic activity in clots of various ages.


RESULTS
The time-related changes in FXIIIa activity in clots was measured with (1) alpha(2)-antiplasmin (alpha(2)AP), a physiological glutamine substrate; (2) alpha(2)AP(13-24), a peptide; and (3) pentylamine, a nonspecific lysine substrate. The cross-linking of alpha(2)AP, alpha(2)AP(13-24), and pentylamine into fibrin by clot-bound FXIIIa declined rapidly with half-lives of 19, 21, and 26 minutes, respectively. Mutational studies showed that glutamine 14 (but not glutamine 3 or 16) and valine 17 of alpha(2)AP(13-24) were required for efficient cross-linking to fibrin. The loss of activity was not due primarily to FXIIIa proteolysis and was partially restored by reducing agents, suggesting that oxidation contributes to the loss of the enzyme's activity in clots. In vivo, the ability of thrombus-bound FXIIIa to cross-link an infused alpha(2)AP(13-24) peptide into existing pulmonary emboli also declined significantly over time.


CONCLUSIONS
FXIIIa cross-links alpha(2)AP and an alpha(2)AP peptide, in a sequence-specific manner, into formed clots with a catalytic half-life of approximately 20 minutes. This indicates that FXIIIa activity is a hallmark of new thrombi and that the antifibrinolytic cross-linking effects of FXIIIa are achieved more rapidly in thrombi than previously believed.
Robinson
B R
BR
Harvard School of Public Health, Harvard Medical School and Massachusetts General Hospital, Boston, MA, 02115, USA.
Houng
A K
AK
Reed
G L
GL
D000375:Aging / Q000502:physiology*; D000818:Animals; D003432:Cross-Linking Reagents / Q000494:pharmacology; D004351:Drug Resistance / Q000502:physiology; D005289:Ferrets; D005337:Fibrin / Q000378:metabolism; D005342:Fibrinolysis / Q000502:physiology*; D005343:Fibrinolytic Agents / Q000494:pharmacology; D006207:Half-Life; D006801:Humans; D007457:Iodine Radioisotopes; D051379:Mice; D008810:Mice, Inbred C57BL; D011655:Pulmonary Embolism / Q000201:enzymology; D013927:Thrombosis / Q000201:enzymology* / Q000503:physiopathology*; D011503:Transglutaminases / Q000378:metabolism*
D003432:Cross-Linking Reagents; D005343:Fibrinolytic Agents; D007457:Iodine Radioisotopes; D005337:Fibrin; D011503:Transglutaminases