Torbet J
Institut Laue-Langevin, Grenoble, France.
Biochem J. 1987 Jun 15;244(3):633-7. doi: 10.1042/bj2440633.
Magnetically induced birefringence was used to monitor fibrin polymerization after the release of the small negatively charged A fibrinopeptides from human fibrinogen by the action of the snake-venom-derived enzymes reptilase and ancrod. A range of conditions was investigated. Fibrin polymerization in solutions of purified fibrinogen shows a distinct break near the gelation point. On addition of Ca2+ or albumin the lag period is shortened, fibre thickness is increased and the break in assembly almost vanishes, probably because both of these additives promote lateral aggregation. There are minor differences in the kinetics, depending on the venom enzyme used. The kinetics of fibrin assembly in model systems containing either Ca2+ or albumin and in human plasma with a largely dormant coagulation cascade are very similar. Therefore in the latter condition there is no significant alteration in the assembly process due to interaction between fibrin or the venom enzymes and any of the plasma proteins. When the cascade is activated, the polymerization progress curves have a character that resembles a combination of the reactions observed when the venom enzymes and endogenously generated thrombin separately induce coagulation, except for a region near gelation where, paradoxically, polymerization appears to be slower on activation. The low-angle neutron-diffraction patterns from oriented gels made with thrombin or reptilase are identical. Therefore at low resolution the packing of the monomers within fibres is the same when fibrinopeptide A only or both fibrinopeptides A and B are removed.
利用磁诱导双折射来监测蛇毒衍生酶类蛇凝血酶和安克洛酶作用于人类纤维蛋白原释放带负电荷的小A纤维蛋白肽后纤维蛋白的聚合过程。研究了一系列条件。纯化纤维蛋白原溶液中的纤维蛋白聚合在凝胶化点附近呈现明显的转折点。添加Ca2+或白蛋白后,延迟期缩短,纤维厚度增加,组装过程中的转折点几乎消失,这可能是因为这两种添加剂都促进了横向聚集。根据所使用的蛇毒酶不同,动力学存在细微差异。在含有Ca2+或白蛋白的模型系统以及凝血级联基本处于休眠状态的人血浆中,纤维蛋白组装的动力学非常相似。因此,在后一种情况下,由于纤维蛋白或蛇毒酶与任何血浆蛋白之间的相互作用,组装过程没有显著改变。当级联被激活时,聚合进程曲线具有一种特征,类似于分别由蛇毒酶和内源性产生的凝血酶诱导凝血时观察到的反应的组合,但在凝胶化附近的一个区域除外,奇怪的是,激活时聚合似乎更慢。用凝血酶或蛇凝血酶制备的取向凝胶的低角度中子衍射图谱是相同的。因此,在低分辨率下,仅去除纤维蛋白肽A或同时去除纤维蛋白肽A和B时,纤维内单体的堆积是相同的。