Department of Chemistry and Applied Biosciences, ETH Zurich, CH-8093, Zurich, Switzerland.
Nat Commun. 2020 Jan 28;11(1):566. doi: 10.1038/s41467-019-14179-x.
Characterizing folding and complex formation of biomolecules provides a view into their thermodynamics, kinetics and folding pathways. Deciphering kinetic intermediates is particularly important because they can often be targeted by drugs. The key advantage of native mass spectrometry over conventional methods that monitor a single observable is its ability to identify and quantify coexisting species. Here, we show the design of a temperature-jump electrospray source for mass spectrometry that allows one to perform fast kinetics experiments (0.16-32 s) at different temperatures (10-90 °C). The setup allows recording of both folding and unfolding kinetics by using temperature jumps from high to low, and low to high, temperatures. Six biological systems, ranging from peptides to proteins to DNA complexes, exemplify the use of this device. Using temperature-dependent experiments, the folding and unfolding of a DNA triplex are studied, providing detailed information on its thermodynamics and kinetics.
生物分子的折叠和复合物形成特性为研究其热力学、动力学和折叠途径提供了重要线索。解析动力学中间态尤为重要，因为这些中间态往往是药物的作用靶点。与传统的仅监测单一可观测变量的方法相比，原位质谱的主要优势在于其能够识别和定量共存的物种。本文介绍了一种用于质谱的温度跳跃电喷雾源的设计，该设计允许在不同温度（10-90°C）下进行快速动力学实验（0.16-32 s）。该装置通过从高温到低温和从低温到高温的温度跳跃，实现了折叠和去折叠动力学的同时记录。六个生物系统，从肽到蛋白质再到 DNA 复合物，都例证了该设备的应用。通过温度依赖性实验，研究了 DNA 三聚体的折叠和去折叠，提供了有关其热力学和动力学的详细信息。
