Department of Cell and Molecular Biology , Northwestern University Feinberg School of Medicine , Chicago , Illinois 60611 , United States.
J Am Chem Soc. 2018 Jul 5;140(26):8060-8063. doi: 10.1021/jacs.8b02918. Epub 2018 Jun 21.
Label-free assays, and particularly those based on the combination of mass spectroscopy with surface chemistries, enable high-throughput experiments of a broad range of reactions. However, these methods can still require the incorporation of functional groups that allow immobilization of reactants and products to surfaces prior to analysis. In this paper, we report a traceless method for attaching molecules to a self-assembled monolayer for matrix-assisted laser desorption and ionization (SAMDI) mass spectrometry. This method uses monolayers that are functionalized with a 3-trifluoromethyl-3-phenyl-diazirine group that liberates nitrogen when irradiated and gives a carbene that inserts into a wide range of bonds to covalently immobilize molecules. Analysis of the monolayer with SAMDI then reveals peaks for each of the adducts formed from molecules in the sample. This method is applied to characterize a P450 drug metabolizing enzyme and to monitor a Suzuki-Miyaura coupling chemical reaction and is important because modification of the substrates with a functional group would alter their activities. This method will be important for high-throughput experiments in many areas, including reaction discovery and optimization.
无标记分析，特别是基于质谱与表面化学相结合的无标记分析，能够实现广泛反应的高通量实验。然而，这些方法仍然需要引入官能团，以便在分析之前将反应物和产物固定在表面上。在本文中，我们报告了一种用于将分子附着到自组装单层上的无痕迹方法，用于基质辅助激光解吸电离（SAMDI）质谱。该方法使用的单层具有 3-三氟甲基-3-苯基重氮基团，当受到照射时会释放氮气，并产生卡宾，插入各种键以共价固定分子。然后，通过 SAMDI 对单层进行分析，可揭示样品中每个分子形成的加合物的峰。该方法用于表征 P450 药物代谢酶，并监测 Suzuki-Miyaura 偶联化学反应，这很重要，因为用官能团修饰底物会改变它们的活性。这种方法将在许多领域的高通量实验中非常重要，包括反应发现和优化。
