Department of Pharmaceutical Sciences, University of Maryland, Baltimore Maryland 21201, United States.
J Am Soc Mass Spectrom. 2021 Jul 7;32(7):1644-1647. doi: 10.1021/jasms.1c00038. Epub 2021 Jun 25.
Fast photochemical oxidation of proteins (FPOP) has demonstrated the ability to inform on the higher order structure of proteins. Recent technological advances have extended FPOP to live cells (IC-FPOP) using multiple cell lines and (IV-FPOP) using . These innovations allow proteins to be studied in their native cellular environment. Hydroxyl radicals are generated via the photoloysis of hydrogen peroxide. Hydrogen peroxide is a signaling molecule that can induce changes to some proteins in the cell limiting the proteins that can be studied by IC-FPOP. Here, we evaluate the sulfate radical anion as a footprinting label in IC-FPOP with sodium persulfate as the precursor. Our findings show a 1.5-fold increase in the number of modified proteins compared to IC-FPOP using hydroxyl radicals at the same precursor concentration demonstrating the amenability of this radical with IC-FPOP.
快速光化学氧化蛋白质（FPOP）已经证明了其能够提供蛋白质高级结构信息的能力。最近的技术进步已经将 FPOP 扩展到了使用多种细胞系的活细胞（IC-FPOP）和使用 的静脉内 FPOP（IV-FPOP）。这些创新使得蛋白质能够在其天然的细胞环境中进行研究。羟基自由基是通过过氧化氢的光解产生的。过氧化氢是一种信号分子，可以诱导细胞中的一些蛋白质发生变化，从而限制了可以通过 IC-FPOP 进行研究的蛋白质。在这里，我们评估了过硫酸氢盐作为前体的 IC-FPOP 中的硫酸根自由基阴离子作为足迹标记。我们的研究结果表明，与使用相同前体浓度的羟基自由基的 IC-FPOP 相比，修饰蛋白的数量增加了 1.5 倍，这表明这种自由基与 IC-FPOP 的适用性。
