Clotting of fibrinogen. 1. Scanning calorimetric study of the effect of calcium.
The denaturation temperature Td and the enthalpy of thermal denaturation delta Hd of the D nodules of fibrinogen increase 12-13 degrees C and 40%, respectively, when fibrinogen is clotted by thrombin in the presence of 10(-3) M calcium ion. The rate of change of Td and delta Hd is first order in thrombin concentration. In the absence of calcium, little change in Td is observed, but the increase in delta Hd still occurs. The shift in Td as a function of logarithm of calcium concentration is sigmoid, with a half-point at 2.5 X 10(-5) M calcium for human and 6.0 X 10(-5) M calcium for bovine fibrinogens, suggesting that the shift is due to binding of calcium at the high-affinity binding sites of fibrin. The Td of the D nodule of native fibrinogen also increases, but not as much, on addition of calcium. This increase in Td is also sigmoid with log calcium, with a half-point of 1.6 X 10(-3) M calcium for human and 3.2 X 10(-3) M calcium for bovine fibrinogens, and appears to be due to binding of calcium to the low-affinity binding sites of fibrinogen. At calcium concentrations greater than 10(-4) M, traces of factor XIII in the bovine fibrinogen preparation become activated and cause cross-linking of the fibrin gel. But the changes in Td and delta Hd still occur when factor XIIIa is inactivated by iodoacetamide, and the rate of the changes is not altered by addition of large amounts of factor XIIIa.(ABSTRACT TRUNCATED AT 250 WORDS)
Donovan
J W
JW
Mihalyi
E
E
D000818:Animals; D001777:Blood Coagulation / Q000187:drug effects*; D002118:Calcium / Q000494:pharmacology*; D002152:Calorimetry, Differential Scanning; D002417:Cattle; D004492:Edetic Acid / Q000494:pharmacology; D005176:Factor XIII / Q000378:metabolism; D005340:Fibrinogen / Q000502:physiology*; D006801:Humans; D007700:Kinetics; D011489:Protein Denaturation; D013816:Thermodynamics; D013917:Thrombin / Q000378:metabolism; D011503:Transglutaminases
D005340:Fibrinogen; D005176:Factor XIII; D004492:Edetic Acid; D011503:Transglutaminases; D013917:Thrombin; D002118:Calcium