Thermodynamics of the cyclophilin-A/cyclosporin-A interaction: a direct comparison of parameters determined by surface plasmon resonance using Biacore T100 and isothermal titration calorimetry.
Thermodynamics of the cyclophilin-A/cyclosporin-A interaction: a direct comparison of parameters determined by surface plasmon resonance using Biacore T100 and isothermal titration calorimetry.
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DOI：
10.1016/j.ab.2006.08.038
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发表时间：
2006-12
影响因子：
2.9
通讯作者：
M. Wear;M. Walkinshaw
中科院分区：
文献类型：
--
作者：
M. Wear;M. Walkinshaw
表面等离子体共振(SPR)1经常用于生物分子相互作用的动力学分析和小分子药物发现/HIT验证研究[1，2]。SPR仪器独一无二地允许测量与络合物形成和解离相关的动力学和热力学数据。此类SPR数据可用于“热力学图谱”的相互作用[3-5]增强溶液结合测量与结构特征的相关性，并将比例能量贡献分配给各个官能团，这是工程疗法和药物设计的基于整体结构的设计方法的基础[6，7]。在这里，我们比较了使用Biacore T100(来自Biacore AB(瑞典乌普萨拉)的一种新的SPR生物传感器)和等温滴定热量计(ITC)(确定平衡热力学的标准方法)的SPR所确定的热力学参数，以了解人亲环素-A(CypA)与其天然抑制剂--疏水性环十一肽环孢素-A(CsA)[9]之间的相互作用。CypA/CsA结构已被解析[10，11]，并很好地表征了相互作用的生物化学[9，12-14]，从而使我们能够合理地获得关于分子机制和结构的热力学数据，并能够更准确地评估和比较从SPR和ITC获得的参数。
Surface plasmon resonance (SPR) 1 is regularly exploited in the kinetic analysis of biomolecular interactions and small-molecule drug-discovery/hit-validation studies [1, 2]. SPR instruments uniquely allow the measurement of kinetic and thermodynamic data specifically associated with complex formation and dissociation. Such SPR data can be used to ‘‘thermodynamic profile’’interactions [3–5] enhancing the correlation of solution binding measurements with structural features and the assignment of proportional energetic contributions to individual functional groups, the basis of the whole-structure-based design approach to engineered therapeutics and drug design [6, 7]. Here we compare thermodynamic parameters determined by SPR using Biacore T100 (a new SPR biosensor from Biacore AB (Uppsala, Sweden)) and by isothermal titration calorimetry (ITC), the standard method of determining equilibrium thermodynamics [8], for the interaction between human cyclophilin-A (CypA) and its natural inhibitor, a hydrophobic cyclic undecapeptide, cyclosporin-A (CsA)[9]. The CypA/CsA structure has been solved [10, 11] and the biochemistry of the interaction well characterized [9, 12–14], thus it allowed us to rationalize the thermodynamic data with regards to the the molecular mechanism and structure and enabled a more accurate assessment and comparison of the parameters obtained from SPR and ITC.