Laboratoire de Biosenseurs & Nanomachines, Département de chimie, Université de Montréal, Montréal, Québec, Canada.
Chemical Computing Group ULC, Montréal, Québec, Canada.
Nat Methods. 2022 Jan;19(1):71-80. doi: 10.1038/s41592-021-01355-5. Epub 2021 Dec 30.
Understanding the relationship between protein structural dynamics and function is crucial for both basic research and biotechnology. However, methods for studying the fast dynamics of structural changes are limited. Here, we introduce fluorescent nanoantennas as a spectroscopic technique to sense and report protein conformational changes through noncovalent dye-protein interactions. Using experiments and molecular simulations, we detect and characterize five distinct conformational states of intestinal alkaline phosphatase, including the transient enzyme-substrate complex. We also explored the universality of the nanoantenna strategy with another model protein, Protein G and its interaction with antibodies, and demonstrated a rapid screening strategy to identify efficient nanoantennas. These versatile nanoantennas can be used with diverse dyes to monitor small and large conformational changes, suggesting that they could be used to characterize diverse protein movements or in high-throughput screening applications.
理解蛋白质结构动态与功能之间的关系，对于基础研究和生物技术都至关重要。然而，用于研究结构快速变化的方法却很有限。在这里，我们引入荧光纳米天线作为一种光谱技术，通过非共价的染料-蛋白质相互作用来感应和报告蛋白质构象变化。我们通过实验和分子模拟，检测并描绘了肠碱性磷酸酶的五个独特构象状态，包括瞬时的酶-底物复合物。我们还利用另一种模型蛋白——Protein G 及其与抗体的相互作用，探索了纳米天线策略的通用性，并展示了一种快速筛选策略，用于鉴定有效的纳米天线。这些多功能的纳米天线可以与各种染料一起使用，以监测小的和大的构象变化，这表明它们可用于描绘各种蛋白质运动，或用于高通量筛选应用。
