Nanoscale Approaches to Understanding Membrane Protein Function

项目摘要/摘要 生物膜是所有生命形式都需要的。这些脂质双层将细胞的内外隔开。 电池，并提供内部隔室之间的分隔。因此，它们是蜂窝网络的站点 识别和交流，并提供了许多蛋白质的家园，这些蛋白质介导信号、催化、 能量的产生和传递以及分子的进出口。尽管这一核心作用在 生命、膜和膜蛋白通常很难用常规的工具来研究 生物化学和分子生物学。膜蛋白在去除后显示活性改变或不活跃 从它们天然的脂肪环境中。同样，揭示了涉及到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。 纳米盘，由一种两亲性支架蛋白溶解的自组装纳米级脂双层 我们的实验室为这些领域的多项新发现提供了服务。在米拉的持续支持下， 我们将利用纳米光盘系统提供新的生化和生物物理途径，以实现 分子机制涉及信号、激素生物合成、药物代谢、外延 低聚病毒抗原和突触连接的膜蛋白的呈递。以下系统 研究包括：人类细胞色素P450系统参与肝脏和肾上腺的代谢；RAS 参与癌症信号传递的GTP酶；纽蛋白，控制细胞迁移的关键成分， 结合寡聚肽的突触连接蛋白，纳米盘排列复合体的能力 低聚表面抗原与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.