The Coagulation of Blood by Snake Venoms and Its Physiologic Significance

Nine of the 17 venoms here tested were found capable of coagulating citrated blood or plasma. As has been believed by most workers in the field, 7 of these 9 coagulant venoms convert fibrinogen to an insoluble modification resembling fibrin (Bothrops atrox, Bothrops jararaca, Bothrops nummifera, Crotalus adamanteus, Crotalus horridus, Crotalus terrificus basiliscus, Crotalus terrificus terrificus). The optimum pH for this coagulation was determined for 3 of these, and was found in each case to be approximately pH 6.5, the same as that for the action of thrombin on fibrinogen. Unlike thrombin, however, the fibrinogen-coagulating activity of the venoms was unaffected by the antithrombin elaborated in the course of anaphylactic shock. In addition to coagulating fibrinogen directly, 3 of these venoms (Bothrops atrox, Bothrops jararaca, and to a less extent, Crotalus terrificus basiliscus) acted on prothrombin to convert it to thrombin, without the necessary intervention of either calcium or platelets. Finally, 2 venoms (Notechis scutatus, and to a slight extent, a mixed Micrurus venom), which had no demonstrable effect on purified fibrinogen, nevertheless converted prothrombin to thrombin. Unlike the reaction between the venoms and fibrinogen, this activation of prothrombin has no definite pH optimum, but takes place over a wide zone (pH 5.6-8.3). In the case of Bothrops atrox, there was some indication that the initial velocity of the reaction increased with increasing alkalinity, but that the amount of thrombin ultimately formed decreased. Extraordinarily minute quantities of some of these venoms sufficed to produce a demonstrable activation of prothrombin. Thus, the fer de lance (Bothrops atrox) venom was active in a 1:25,000,000 dilution, and that of the Australian tiger snake (Notechis scutatus) was active in a 1:4,000,000 dilution. The thrombin formed was indistinguishable from that produced by the action of calcium + platelets on prothrombin. Like the latter type of thrombin, and unlike venoms which act directly on fibrinogen, thrombin formed from prothrombin by venom was inhibited by antithrombin. Every one of the 9 non-coagulant venoms in this series destroyed prothrombin; and 5 of these destroyed fibrinogen as well. As is discussed in the text, there is reason to believe that these several properties of the venoms (coagulation and destruction of fibrinogen; activation and destruction of prothrombin) depend on the proteolytic enzymes which they were found to contain. These observations lend further support to the thesis that, in the course of physiological coagulation, (a) calcium plus platelets (or tissue derivative) constitute an enzyme system which reacts with prothrombin to form thrombin, and which is thus analogous to trypsin and to several of the proteolytic venoms here discussed, and (b) the thrombin so formed is itself a proteolytic enzyme which, like papain and the majority of the coagulant and proteolytic snake venoms here studied, reacts with fibrinogen to form a fibrillar gel, fibrin.