Effects of Clotting on the Label in Iodinated Fibrinogen in Different Species

Summary

1. The behaviour of the iodine label during coagulation was investigated in 8 species using ¹²⁵I-fibrinogen.

2. Guinea pig, human, monkey, ox and rabbit fibrinogen preserved the label quantitatively when converted into fibrin, but goat and sheep fibrinogen lost 5.6% and canine fibrinogen lost 11.1 % of its radioactivity.

3. Using a constant iodination technique, the fraction of radioactivity released during clotting proved to be fairly reproducible. The origin and quantity of thrombin used had no influence on the results.

4. The released activity was tentatively identified in the clot liquor of sheep as ¹²⁵I-fibrinopeptide B. Based on this observation, a theory is presented which satisfactorily explains the experimental findings by the variations in the occurrence of non-esterified tyrosine residues in the fibrinopeptides of different species.

• References

• 1 Bailey K, Bettelheim F. R, Lóránd L, Middlebrook W. R. Action of thrombin in the clotting of fibrinogen. Nature (Lond.) 167: 233-234 1951;
• 2 Bettelheim F. R, Bailey K. The products of the action of thrombin on fibrinogen. Biochim. biophys. Acta (Amst.) 9: 578-579 1952;
• 3 Blombäck B, Blombäck M, Edman P, Hessel B. Human fibrinopeptides. Isolation, characterization and structure. Biochim. biophys. Acta (Amst.) 115: 371-396 1966;
• 4 Blombäck B, Doolittle R. F. Amino acid sequence studies on fibrinopeptides from several species. Acta chem. scand 17: 1819-1822 1963;
• 5 Blombäck B, Doolittle R. F, Blombäck M. Fibrinogen: Structure and evolution. Protides of the Biological Fluids, Proceedings of the 12th Colloquium, Bruges, 1964 Elsevier; Amsterdam: 1965: 87.
• 6 Blombäck B, Sjöquist J. Studies on fibrinopeptides from different species. Acta chem. scand 24: 493-495 1960;
• 7 Caspary E. A, Kekwick R. A. Some physiochemical properties of human fibrinogen. Biochem. J 67: 41-48 1957;
• 8 Doolittle R. F, Blombäck B. Amino acid sequence investigations of fibrinopeptides from various mammals: evolutionary implications. Nature (Lond.) 202: 147-152 1964;
• 9 Gladner J. A, Folk J. E, Laki K, Carroll W. R. Thrombin-induced formation of co-fibrin I and II. J. biol. Chem 234: 62-70 1959;
• 10 Gross D. An apparatus high-voltage paper electrophoresis. J. Chromatog 5: 194-206 1961;
• 11 Haid A. Statistical Tables and Formulas. Wiley & Sons; London: 1952
• 12 Laki K. The action of thrombin on fibrinogen. Science 114: 435-436 1951;
• 13 Laki K. The polymerization of proteins: the action of thrombin on fibrinogen. Arch. Biochem 32: 317-324 1951;
• 14 Laki K. The clotting of fibrinogen. Sci. Amer 206: 60-66 1962;
• 15 Laki K. Enzymatic effects of thrombin. Fed. Proc 24: 794-799 1965;
• 16 Laki K, Gladner J. A. Chemistry and physiology of the fibrinogen-fibrin transition. Physiol, Rev 44: 127-160 1964;
• 17 Loevinger R, Berman M. Efficiency criteria in radioactivity counting. Nucleonics 9: 26-39 1951;
• 18 Lóránd L. ‘Fibrino-peptide’: New aspects of the fibrinogen-fibrin transformation. Nature (Lond.) 167: 992-993 1951;
• 19 Lóránd L. Fibrino-peptide. Biochem. J 52: 200-203 1952;
• 20 Lóránd L, Middlebrook W. R. The action of thrombin on fibrinogen. Biochem. J 52: 196-199 1952;
• 21 Lóránd L, Middlebrook W. R. Studies on fibrino-peptide. Biochim. biophys. Acta (Amst.) 9: 581-582 1952;
• 22 McFarlane A. S. Efficient trace-labelling of proteins with iodine. Nature (Lond.) 182: 53 1958;
• 23 McFarlane A. S. In vivo behaviour of I¹³¹-fibrinogen. J. clin. Invest 42: 346-361 1963;
• 24 McFarlane A. S. Metabolism of Plasma Proteins. In: Munro-Allison: Mammalian Protein Metabolism. Vol. I 333 Academic Press; New York: 1963
• 25 McFarlane A. S, Todd D, Cromwell S. Fibrinogen catabolism in humans. Clin. Sci 26: 415-420 1964;
• 26 Mihályi E, Godfrey J. E. Digestion of fibrinogen by trypsin. I. Kinetic studies of the reaction. Biochim. biophys. Acta (Amst.) 67: 73-89 1963;
• 27 Mihályi E, Godfrey J. E. Digestion of fibrinogen by trypsin. II. Characterization of the large fragment obtained. Biochim. biophys. Acta (Amst.) 67: 90-103 1963;
• 28 Osbahr Jr. A. J, Gladner J. A, Laki K. The action of human thrombin on human fibrinogen. Biochem. biophys. Res. Commun 13: 462-465 1963;
• 29 Oncley J. L. Interpretation of data obtained in studies with isotope-labeled proteins of biological significance. Chemical considerations. Fed. Proc 16 (Suppl. 01) 35 1957;
• 30 Shulman S. The size and shape of bovine fibrinogen. Studies of sedimentation, diffusion and viscosity. J. Amer. chem. Soc 75: 5846-5852 1953;
• 31 Tinoco I, Ferry J. D. The site of attack by thrombin on fibrinogen. J. Amer. chem. Soc 76: 5573-5574 1954;
• 32 Weber E. Grundriß der biologischen Statistik. S. 254. Gustav Fischer Verlag; Jena: 1957