Structural biology and high resolution studies of P-glycoprotein

项目摘要/摘要 小鼠P-糖蛋白(Pgp)的最新X射线结构测定为3.8° 建立了研究最多的哺乳动物之一的整体结构架构 多药耐药性(MDR)转运体。然而，PGP的结构只是一个 为了解其转运机制和潜在的抑制作用奠定了基础。显然 具有不同构象和共晶结构的高分辨率结构 将需要了解与ATP结合的运输的详细机制 水解为未来合理的药物设计铺平了道路。我们建议(1)延长 新型膜蛋白结晶法拆分PGP晶体 技术和新型洗涤剂，大大提高了模型的精度，(2) 测定PgP在内向和向外的额外构象 用于绘制运输周期的结构轨迹的构象，以及(3) 确定两种经典和临床重要的Pgp的共晶结构 阿霉素和罗丹明类似物，以了解多特异性底物 有约束力的。

Project Summary/Abstract The recent x-ray structure of mouse P-glycoprotein (Pgp) determined to 3.8 ¿ established the overall structural architecture of one of the most studied mammalian multidrug resistance (MDR) transporters. The structure of Pgp, however, is only a starting point for understanding its transport mechanism and potential inhibition. Clearly a higher resolution structure along with different conformations and co-crystal structures will be required to understand the detailed mechanism of transport coupled to ATP hydrolysis paving the way towards future rational drug design. We propose to (1) extend the resolution of Pgp crystals using very new membrane protein crystallization techniques and novel detergents to greatly improve the precision of the model, (2) determine additional conformations of Pgp both in the inward- and outward- facing conformations to map out the structural trajectories of the transport cycle, and (3) determine the co-crystal structures of Pgp of two classical and clinically important compounds, doxorubicin and rhodamine analogs, to understand poly-specific substrate binding.