人促红细胞生成素小肽模拟物研究

发布时间：2018-04-22 18:23

本文选题：促红细胞生成素 + 分子动力学模拟　；参考：《吉林大学》2006年博士论文

【摘要】： 本文以分子动力学模拟方法和分子力学理论为基础,构建了自有的人促红细胞生成素(Erythropoietin,EPO)模拟肽(Erythropoietin Mimetic peptide,EMP)与其受体(Erythropoietin Receptor,EpoR)结合的动力学模型。通过对该模型的结构研究和理论分析,对EMP与EpoR结合机制提出了新的理论解释。以该理论为指导,设计合成了新的一系列EPO模拟肽(Mimetic Binding Peptides,MBPs)。经过对MBPs活性的实验检验,验证了理论和模型的准确性,并得到了具有较好活性的MBPs分子。以人EpoR与EPO、EMP的复合物X-光衍射晶体结构(PDB ID:1CN4,1EBP等)为模板,利用该复合结构的X-光衍射结构分析数据,用分子力学和分子动力学方法,建立了EMP与EpoR结合的分子动力学模型,并对这个模型中的配体与受体相互作用机制进行了理论研究。在对模型的结构研究和模拟计算中发现,静电相互作用是配体与受体相互作用的主要作用力。不同配体分子的空间结构和与受体的不同结合方式,会直接影响二聚后两EPO受体分子间夹角和距离,进而产生不同的活性作用——激活剂或者拮抗剂。本文在对此模型的结构分析和配体与受体结合机制研究的理论基础上,设计了一系列新的EPO小肽类模拟物,我们将其命名为MBPs。使用电荷和疏水性质更强的部分氨基酸来取代原有的EMP中的相应氨基酸。经过计算分析,这些新的模拟肽MBPs在理论上应具有更好的结合和激活活性。为了从实验上验证设计后的MBPs的结合与聚合活性,我们利用荧光标记法以及微孔板荧光技术建立了EPO模拟物的体外结合与聚合活性检测方法。由活性检测所得到的实验结果,与理论计算和设计相比较,验证了我们建立的模型的可靠性及计算方法的合理性。为小肽类药物设计和开发开拓了一个新的设计思路和实践方法。
[Abstract]:Based on the method of molecular dynamics simulation and the theory of molecular mechanics, a kinetic model of the binding of human erythropoietin Erythropoietin (Erythropoietin Mimetic peptidea) with its receptor Erythropoietin (EpoR) has been established in this paper. Through the research and theoretical analysis of the structure of the model, a new theoretical explanation of the mechanism of EMP and EpoR is put forward. Under the guidance of this theory, a series of EPO mimic peptides, Mimetic Binding Peptides, were designed and synthesized. The accuracy of the theory and model has been verified by the experimental test of MBPs activity, and the MBPs molecules with good activity have been obtained. Using the complex of EpoR and EPO-EMP as template, using the X-ray diffraction data of EMP ID1 CN41EBP etc., the molecular dynamics model of EMP combined with EpoR was established by molecular mechanics and molecular dynamics method. The mechanism of ligand-receptor interaction in this model was studied theoretically. It is found that electrostatic interaction is the main interaction force between ligand and receptor. The spatial structure of different ligands and their different binding modes with receptors will directly affect the angle and distance between the two EPO receptor molecules after dimerization, and then produce different active activators or antagonists. Based on the theoretical analysis of the structure of the model and the mechanism of ligand binding to receptors, a series of new mimics of small peptides of EPO have been designed and named MBPs. The corresponding amino acids in the original EMP were replaced by some amino acids with stronger charge and hydrophobic properties. By calculation and analysis, these new mimic peptides MBPs should have better binding and activation activity in theory. In order to verify the binding and polymerization activity of the designed MBPs in experiments, we developed an in vitro method for the detection of the binding and polymerization activity of EPO mimics by fluorescence labeling method and microporous plate fluorescence technique. Compared with the theoretical calculation and design, the experimental results obtained from the activity test verify the reliability of the model and the reasonableness of the calculation method. It opens up a new design idea and practical method for the design and development of small peptide drugs.
【学位授予单位】：吉林大学
【学位级别】：博士
【学位授予年份】：2006
【分类号】：R341

【相似文献】