DP9柿子单宁对其靶标蛋白蛇毒PLA2的特异识别与共价修饰位点研究

对柿子特征成分柿单宁个性特征及其与特定靶标蛋白质独特的识别机制与作用途径进行深入研究可为我们从分子水平阐释其特殊药理作用提供了一条新思路。我们在前期研究中首次发现具有特殊结构的DP9柿单宁体内外解蛇毒功效最为显著，特别是发现其对蛇毒蛋白中磷脂酶A2有特殊的亲合力，能通过共价作用不可逆改变其靶标蛇毒PLA2的结构和功能，但DP9柿子单宁对蛇毒PLA2特异性识别途径及共价作用位点尚不清楚。本项目将在前期实验结果及首次发现DP9柿子单宁能共价修饰蛇毒PLA2结构的基础上采用SPR、MALDI-TOF-MS、固态NMR、分子模拟及生物学信息等多种新技术结合继续和进一步深入研究DP9柿子单宁对其靶标蛇毒PLA2的特异性识别模式及具体的共价交联位点。项目的顺利完成将为阐述柿单宁特殊药学作用的分子机制提供新理论，为解蛇毒药物的研制提供新靶点，也将为研究复杂多酚类化合物与蛋白质的相互作用提供方法参考。

Carrying on a further research on the unique structure of persimmon tannin and the specific recognition mechanism of persimmon tannin on its target proteins may provide a novel pathway from the molecular level to explain its special pharmacological effects. In our previous studies we found for the first time that DP9 persimmon tannin had a special structure and was the most effective detoxifying component against snake venom both in vitro and in vivo, especially it had a special affinity on snake venom phospholipase A2 (PLA2) , and could modify its structure covalently, thus irreversibly changed the structure and function of PLA2. But the detailed recognition pathway and the specific covalent binding sites remain unclear. Based on the previous results and the first finding of covalent modification of PLA2 by DP9 persimmon tannin, this project will further study the specific recognition mode and detailed covalent binding sites of DP9 persimmon tannin on its target protein PLA2 by combining use of SPR, MALDI-TOF-MS, solid NMR, molecular modeling and other new technology. The successful completion of this project will help develop new strategies for understanding the special pharmaceutical activity of DP9 persimmon tannin, and will provide novel targets for developing anti-venom drugs, and will supply reference methods for studying the interaction of complex polyphenols with protein as well.