Deciphering the relationship between structure, dynamics and function in helical bundle proteins

项目摘要/摘要 该方案结合了病毒离子通道和从头蛋白设计。我们在M2方面的工作 甲型流感病毒的质子通道主要通过抑制和质子运动的机制来实现 频道。M2也是金刚烷胺类流感药物的靶标，而且大多数甲型流感病毒分离株 病毒现在对金刚烷胺具有抗药性。将使用结晶学、计算和2DIR来询问 抗金刚烷胺病毒株中抑制M2的药物的机制和辅助设计。同时， 我们将扩大我们的研究范围，以探讨E蛋白的传导和抑制机制。 冠状病毒，其结构和功能与M2基本相似。 从头开始的蛋白质设计提供了一种测试和改进我们对蛋白质结构和 功能。我们正在开发计算方法来设计与小分子结合的蛋白质，例如 药物和金属卟啉。我们还在设计膜蛋白，以阐明 它们可以折叠并发挥作用。为了探索高选择性质子传导蛋白的机制，我们正在 设计质子选择性通道，以检验水分子网络(水线)可以 通过蛋白质中质子传导途径的非极延伸动态地和瞬时地形成。这个 水线将允许质子传导，但不能传导K+或Na+等大的水合碱金属离子。我们 也参与了蛋白质的设计，这些蛋白质结合并运输氨基酸等营养物质。 膜，并设计屏幕在体内测试它们。

Project Summary/Abstract This proposal combines work on viral ion channels and de novo protein design. Our work on the M2 proton channel from influenza A virus focuses on the mechanism of inhibition and proton movement through the channel. M2 is also the target of the amantadine class of influenza drugs, and most isolates of influenza A virus are now amantadine-resistant. Crystallography, computation and 2DIR will be used to interrogate the mechanism and aid in design of drugs that inhibit M2 in amantadine-resistant strains of the virus. In parallel, we will expand our studies to examine the mechanism of conduction and inhibition of the E-proteins from coronaviruses, which have structures and functions largely similar to M2. De novo protein design provides a means to test and refine our understanding of protein structure and function. We are developing computational methods to design proteins that bind small molecules such as drugs and metalloporphyrins. We are also designing membrane proteins to elucidate the principles by which they fold and function. To probe the mechanisms of highly selective proton conduction proteins, we are designing proton-selective channels that test a hypothesis that networks of water molecules (water wires) can form dynamically and transiently through apolar stretches of the proton conduction pathway in proteins. The water wires would allow conduction of protons but not large hydrated alkali metal ions such as K+ or Na+. We also are engaged in design of proteins that bind to and transport nutrients such as amino acids across membranes, and devising screens to test them in vivo.