毛细管区带电泳技术研究龙血竭有效单体与核酸适配体及凝血酶之间的相互作用

Interactions Between Aptamer, Bovine Thrombin and Active Molecules in Dragon's Blood by CZE

  • 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Kb为4.73×104 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Kb分别为1.98×104,1.83×104 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.

     

    Abstract: 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 Kb between Loureirin A and B-Thr is 4.73×104 L/mol, and Loureirin B, Loureirin C, Resveratrol 7,4-dihydroxyflavanone are no bound with B-Thr obviously. The Kb between Loureirin B, DHF and Apt29 are 1.98×104, 1.83×104 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.

     

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