Nanoscale Approaches to Understanding Membrane Protein Function

项目总结/摘要 生物膜是所有生命形式所必需的。这些脂质双层将细胞的内部和外部分开， 电池并提供内部隔室之间的分隔。因此，它们是蜂窝网络 识别和通信，并为介导信号传导，催化， 能量的产生和传递以及分子的输入和输出。尽管这一核心作用， 生命、膜和膜蛋白通常很难用常规的生物学工具来研究。 生物化学和分子生物学。膜蛋白显示改变的活性或被去除时无活性 从它们的天然脂质环境中分离出来同样地，揭示涉及的基本分子识别事件 在膜表面形成复杂的多组分结构需要新的方法。 纳米圆盘，自组装的纳米级脂质双层溶解的两亲性支架蛋白， 我们的实验室，已经在这些领域做出了多项新的发现。在MIRA的持续支持下， 我们将使用Nanodisc系统提供新的生物化学和生物物理路径，以实现 参与信号传导的分子机制，激素生物合成，药物代谢， 寡聚病毒抗原和突触连接的膜蛋白的呈递。系统在 研究包括：人细胞色素P450系统涉及肝脏和肾上腺代谢; Ras 参与癌症信号传导的GTPases;粘着斑蛋白是控制细胞迁移的关键成分， 结合寡聚肽的突触连接蛋白，纳米盘有序复合物的能力 寡聚体表面抗原和纳米盘组装的生物物理学。

Project Summary / Abstract Biological membranes are needed by all life forms. These lipid bilayers separate the inside and outside of the cell and provide the separation between internal compartments. As such, they are the site for cellular recognition and communication and provide the home to a host of proteins that mediate signaling, catalysis, the generation and transduction of energy and the import and export of molecules. Despite this central role in life, membranes and membrane proteins have often been difficult to study using the normal tools of biochemistry and molecular biology. Membrane proteins display altered activity or are inactive when removed from their native lipid environment. Likewise, revealing the fundamental molecular recognition events involved in forming complex multi-component architectures at the membrane surface requires new methodologies. Nanodiscs, self-assembled nanoscale lipid bilayers solubilized by an amphipathic scaffold protein developed in our laboratory, have served to enable multiple new discoveries in these arenas. Under continued MIRA support, we will use the Nanodisc system to provide novel biochemical and biophysical paths to the realization of the molecular mechanisms involved in signaling, hormone biosynthesis, drug metabolism, the epitaxial presentation of oligomeric viral antigens and the membrane proteins of the synaptic junction. Systems under investigation include: The human cytochrome P450 systems involved liver and adrenal metabolism; the Ras GTPases involved in cancer signaling; vinculin, a critical component in the control of cellular migration, the proteins of the synaptic junction that bind oligomeric peptides, the ability of Nanodiscs to order complex oligomeric surface antigens and the biophysics of Nanodisc assembly.