Developing next-generation nanodiscs for the study and modulation of membrane proteins

摘要 膜蛋白是支持细胞生长、介导信号转导和细胞增殖的重要分子。 保持体内平衡它们要么与脂质双层结合，要么嵌入脂质双层，因此表现出 复杂的化学性质和生物物理研究提出了艰巨的挑战。过去 几十年来，巨大的努力集中在提供这些复杂的机械见解， 大分子然而，由于缺乏强有力的工具来促进结构调整， 和膜蛋白的功能研究。此外，水不溶性脂质双层使其 为了治疗目的调节这些蛋白质是非常困难的。为了解决这些问题， 拟议的研究旨在开发下一代纳米盘， 简单的询问和调节膜蛋白。首先，我们将开发一种清洁剂- 自由纳米盘系统，以便于从其天然脂质中分离完整的膜蛋白 环境第二，我们将设计纳米盘报告器，以允许快速和无标记检测 外周膜蛋白的膜结合和重塑活性。最后，我们将建立 具有前所未有的特异性的融合和可转换的纳米盘到药物膜蛋白， 功效总之，这些简单而多功能的工具将大大改变膜领域 生物化学和揭示了膜蛋白，以前被认为是新的光 不可接近的或者不可拒绝的。

Abstract Membranes proteins are essential molecules to support cell growth, mediate signal transduction, and maintain homeostasis. They are either associated with, or embedded in the lipid bilayer, thus exhibit complex chemical properties and present formidable challenges for biophysical studies. Over the past few decades, enormous efforts have focused on providing mechanistic insights into these sophisticated macromolecules. However, the progress is hampered by the lack of robust tools to facilitate structural and functional studies for membrane proteins. Moreover, the water-insoluble lipid bilayer makes it notoriously difficult to modulate these proteins for therapeutic purposes. To address these problems, the proposed research aims to develop next-generation nanodiscs that enact simple and straightforward interrogation and modulation of membrane proteins. First, we will develop a detergent- free nanodisc system to ease the isolation of integral membrane proteins from their native lipid environment. Second, we will engineer nanodisc reporters to allow rapid and label-free detection of membrane binding and remodeling activities of peripheral membrane proteins. Finally, we will build fusogenic and switchable nanodiscs to drug membrane proteins with unprecedented specificity and efficacy. Together, these simple and versatile tools will significantly transform the field of membrane biochemistry and shed new light on membrane proteins that were previously thought to be unapproachable or undruggable.