Abstract
Clots of human beta-fibrin, in which only (or predominantly) the B fibrinopeptide is released, were formed at 14 degrees C by copperhead venom procoagulant enzyme (CVE or venzyme), at pH 8.5, ionic strength 0.45. The shear modulus of elasticity increased slowly and after several days attained a constant value, which was lower than those of alpha-fibrin or alpha beta-fibrin under the same conditions. Before studying the temperature dependence of elasticity, the CVE was then inhibited by introducing phenyl methyl sulfonyl chloride (PMSF) by diffusion. With increasing temperature, the modulus decreased progressively from 5 degrees C to nearly zero at 35 degrees and was essentially reversible with temperature change; recovery of elasticity after change from 34.5 degrees to 14 degrees required approximately 2 d but was considerably faster than the initial buildup of elasticity by CVE at 14 degrees. Creep and creep recovery measurements on unligated clots showed creep rates and irrecoverable deformation that were similar in magnitude to those of alpha-fibrin clots formed with batroxobin and much larger than those of alpha beta-fibrin clots formed with thrombin, under the same conditions. During creep and creep recovery, the differential modulus or compliance remained constant, showing that there was no permanent structural damage, and if network strands are severed in slow flow, they must rejoin in new configurations. Introduction (by diffusion) of the tetrapeptides Gly-His-Arg-Pro (GHRP) and Gly-Pro-Arg-Pro (GPRP), which resemble the B and A binding sites on the E domain of fibrin respectively, reduced the shear modulus and increased the creep rate of beta-fibrin clots to an extent similar to the effect of GPRP on alpha beta-fibrin, much more than that of GHRP on alpha beta-fibrin, but much less than that of GPRP on a-fibrin. A ligated beta-fibrin clot formed with Factor XIIIa (in which the activating thrombin had been neutralized by hirudin) showed essentially perfect elastic behavior, with no creep and with complete recovery after removal of stress, and was inert to GHRP.