Harris G, Doctor V M
Chemistry Department of Prairie View A University, Texas 77446, USA.
Blood Coagul Fibrinolysis. 2002 Jun;13(4):355-9. doi: 10.1097/00001721-200206000-00012.
Studies were conducted on the effect of 6-aminohexanoic acid (6-AH) or fucoidan on the activation of glutamic plasminogen (glu-plg) by streptokinase using 0.05 mol/l Tris buffer containing a physiological concentration of NaCl. In contrast to the earlier reports where no NaCl was added to the buffer solution, addition of 6-AH enhanced the initial rate while the inhibition by fucoidan was not affected. Double reciprocal plots of the activation of glu-plg by streptokinase in the presence of 6-AH showed an increase in Vmax, but no change in Km. However, the addition of fucoidan showed a decrease in Vmax, but no change in Km. To determine whether the stimulatory effect of 6-AH was specifically directed towards glu-plg or streptokinase, the ratios of the initial rate of plasmin generation in the presence of 6-AH over the controls were plotted against the inverse of the volume fraction of glu-plg or streptokinase after serial dilutions. The results indicated that the dilutions of glu-plg, but not of streptokinase, influenced the ratios, suggesting an interaction of 6-AH with glu-plg. Similar experiments were conducted to determine the mechanism of inhibition of streptokinase by fucoidan. The results indicated that fucoidan was interacting with streptokinase, but not with glu-plg. Circular dichroism studies of glu-plg in the near-ultraviolet spectra (250-308 nm) showed that addition of 6-AH enhanced the spectra in the region around certain chromophores, which reflected conformational changes. On the contrary, the far-ultraviolet spectra were almost identical.
使用含有生理浓度氯化钠的0.05 mol/l Tris缓冲液,研究了6-氨基己酸(6-AH)或岩藻依聚糖对链激酶激活谷氨酸纤溶酶原(glu-plg)的影响。与早期报道中缓冲溶液未添加氯化钠不同,添加6-AH可提高初始速率,而岩藻依聚糖的抑制作用不受影响。在存在6-AH的情况下,链激酶激活glu-plg的双倒数图显示Vmax增加,但Km不变。然而,添加岩藻依聚糖显示Vmax降低,但Km不变。为了确定6-AH的刺激作用是特异性针对glu-plg还是链激酶,将存在6-AH时纤溶酶生成的初始速率与对照的比率,针对连续稀释后glu-plg或链激酶体积分数的倒数作图。结果表明,glu-plg的稀释而非链激酶的稀释影响该比率,表明6-AH与glu-plg相互作用。进行了类似实验以确定岩藻依聚糖抑制链激酶的机制。结果表明,岩藻依聚糖与链激酶相互作用,但不与glu-plg相互作用。glu-plg在近紫外光谱(250-308 nm)的圆二色性研究表明,添加6-AH增强了某些发色团周围区域的光谱,这反映了构象变化。相反,远紫外光谱几乎相同。