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 保持动态平衡。它们要么与脂双层结合，要么嵌入脂双层中，从而显示 复杂的化学性质，对生物物理研究提出了严峻的挑战。在过去的时间里 几十年来，巨大的努力集中在为这些复杂的 大分子。然而，由于缺乏有力的工具来促进结构性改革，这一进展受到阻碍。 以及膜蛋白的功能研究。此外，水不溶的脂双层使其 为治疗目的调节这些蛋白质是出了名的困难。为了解决这些问题， 这项拟议的研究旨在开发下一代纳米光盘，这种光盘可以制作简单和 直截了当地询问和调节膜蛋白。首先，我们将开发一种洗涤剂-- 用于简化从天然脂中分离完整膜蛋白的自由纳米盘系统 环境。其次，我们将改造纳米盘记者，使其能够快速和无标签地检测到 外周膜蛋白的膜结合和重塑活性。最后，我们将建立 融合和可切换的纳米盘对药物膜蛋白具有前所未有的特异性和 功效。这些简单而通用的工具将极大地改变薄膜领域 生物化学，并揭示了以前被认为是 无法接近或无法下药的。

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.