Autoprothrombin C Activity from Prothrombin with Snake Venom or Trypsin^[*]

Summary

Suitable conditions were found for the generation of autoprothrombin C from purified prothrombin with the use of Russell’s viper venom or trypsin. DEAE chromatographed prothrombin is structurally altered and has never been found to yield autoprothrombin C and also did not yield it when Russell’s viper venom or trypsin were used. Autoprothrombin C is derived from prothrombin with tissue extract thromboplastin, but not in large amounts with the intrinsic clotting factors. With the latter thrombin and autoprothrombin III are the chief activation products. Autoprothrombin III concentrates were prepared from serum and upon activation with 25% sodium citrate solution or with Russell’s viper venom large amounts of autoprothrombin C were obtained, and this was of high specific activity. Theoretically trypsin is not a thrombolytic agent, but on the contrary should lead to intravascular clotting.

^* This investigation was supported by a research grant HE-05141 from the National Heart Institute, National Institutes of Health, U. S. Public Health Service,

^** Rockefeller Foundation Research Fellow.

• 1 Marciniak E, Seegers W. H. Autoprothrombin C: A Second Enzyme from Prothrombin. Canad. J. Biochem. 1962; 40: 597
• 2 Seegers W. H, Cole E. R, Marciniak E. Partial Activation of Purified Prothrombin: Derivation of Autoprothrombin I with Use of Autoprothrombin C. Throm-bos. Diathes. haemorrh. 1962; 7: 239
• 3 Seegers W, Aoki H. N, Marciniak E. Autoprothrombin C and the Generation of Thrombin from Prothrombin Activation Mixtures. New Istanbul Contribution to Clinical Science 1962; 5: 170
• 4 Marciniak E, Cole E. R, Seegers W. H. Functional Alteration in the Prothrombin Molecule. Nature (London) 1962; 195: 1305
• 5 Thomas W. R, Seegers W. H. Terminal Amino Acids of Bovine Prothrombin and Thrombin Preparations. Biochim. biophys. Acta 1960; 42: 556
• 6 Seegers W. H, Marciniak E. Autoprothrombin C in Irregular Blood Clotting. Thrombos. Diathes. haemorrh. 1962; 8: 1
• 7 Landaburu R. H, Barnhart M. I, Seegers W. H. Prothrombin Activation with Trypsin as Enzyme. Amer. J. Physiol. 1961; 201: 298
• 8 Lee C. Y, Johnson S. A, Seegers W. H. Clotting of Blood with Russell's Viper Venom. J. Mich. med. Soc. 1955; 54: 801
• 9 Lee P. H, Johnson S. A, Seegers W. H. Platelet Cothromboplastin: A Platelet Factor(s) Related to the Blood Clotting Mechanisms. Thrombos. Diathes. haemorrh. 1957; 1: 16
• 10 Esnouf M. P, Williams W. J. Purification. Properties and Activation of Factor X. Thrombos. Diathes. haemorrh. 1962; 7 (Suppl.) 213
• 11 Seegers W. H. The Purification of Prothrombin. Rec. Chem. Progr. 1952; 13: 143
• 12 Seegers W. H, Landaburu R. H. Purification of Prothrombin and Thrombin by Chromatography on Cellulose. Canad. J. Biochem. 1960; 38: 1405
• 13 Ware A. G, Seegers W. H. Two-stage Procedure for the Quantitative Determination of Prothrombin Concentration. Amer. J. clin. Path. 1949; 19: 471
• 14 Seegers W. H, Smith H. P. Factors Which Influence the Activity of Purified Thrombin. Amer. J. Physiol. 1942; 137: 348
• 15 Ware A. G, Seegers W. H. Plasma Accelerator Globulin: Partial Purification, Quantitative Determination, and Properties.. J. biol. Chem. 1948; 172: 699
• 16 Hecht E. R, Cho M. H, Seegers W. H. Thromboplastin: Nomenclature and Preparation of Protein-free Material Different from Platelet Factor 3 or Lipid Activator. Amer. J. Physiol. 1958; 193: 584
• 17 McClaughry R. I. Concentration of a Prothrombin Conversion Accelerator and Thrombin Precursor from Serum. Amer. J. Physiol. 1956; 186: 335
• 18 Weiner M. Pharmacological Considerations of Antithrombotic Therapy. Advances in Pharmacology Academic Press; New York and London: 1962. 1 277
• 19 Eagle H. Recent Advances in Blood Coagulation Problem. Medicine (Baltimore) 1937; 16: 95