摘要
利用毛细管区带电泳技术(CZE)研究5个龙血竭有效单体与牛凝血酶(B-Thr)、29碱基凝血酶核酸适配体(Apt29)之间的相互作用.电泳条件为有效长度是40 cm的非涂层石英毛细管,气压进样3.45 kPa,进样时间5 s,分离电压+15 kV,分离温度15℃,检测波长分别为214 nm(牛凝血酶体系)和330 nm(核酸适配体体系).结果表明,龙血素A和B-Thr之间的结合常数K_b为4.73×10~4 L/mol,白藜芦醇、龙血素B、7,4′-二羟基黄酮、龙血素C与B-Thr没有显著结合;龙血素B和7,4′-二羟基黄酮与核酸适配体的结合常数K_b分别为1.98×10~4,1.83×10~4 L/mol,白藜芦醇、龙血素A、龙血素C与核酸适配体之间没有表现出明显的结合现象.本文结果对进一步研究龙血竭在凝血酶上的潜在结合位点,以及核酸适配体能否向凝血酶靶向转运龙血竭,提供了科学数据支持.
Interactions between Apt29, bovine thrombin(B-Thr) and five active molecules in Dragon's blood were investigated based on capillary zone electrophoresis(CZE). The CZE conditions were set as following: effective capillary length of 40 cm, injection volume of 3.45 kPa, injection time of 5 s, voltage of +15 kV, temperature of 15 ℃, detection wavelength of 214 nm and 330 nm. The results show that K_b between Loureirin A and B-Thr is 4.73×10~4 L/mol, and Loureirin B, Loureirin C, Resveratrol 7,4-dihydroxyflavanone are no bound with B-Thr obviously. The K_b between Loureirin B, DHF and Apt29 are 1.98×10~4, 1.83×10~4 L/mol, respectively. Loureirin A, Loureirin C and Resveratrol are no bound with Apt29 obviously. The study indicates the interactions between B-Thr, Apt29 and active molecules in Dragon's blood. It can provide a basis for speculating the potential binding site of dragon's blood with thrombin, and delivering Dragon's blood to thrombin with aptamer as target delivery tool.
引文
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