Chemical Penetration Enhancers Increase Hydrogen Peroxide Uptake in C. elegans for In Vivo Fast Photochemical Oxidation of Proteins.,Journal of Proteome Research

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Chemical Penetration Enhancers Increase Hydrogen Peroxide Uptake in C. elegans for In Vivo Fast Photochemical Oxidation of Proteins.
Journal of Proteome Research ( IF 3.8 ) Pub Date : 2020-06-07 , DOI: 10.1021/acs.jproteome.0c00245
Jessica A Espino ₁ , Zhihui Zhang ₁ , Lisa M Jones ₁

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Fast photochemical oxidation of proteins (FPOP) is a hydroxyl radical protein footprinting method that covalently labels solvent-accessible amino acids by photolysis of hydrogen peroxide. Recently, we expanded the use of FPOP for in vivo (IV-FPOP) covalent labeling in C. elegans. In initial IV-FPOP studies, 545 proteins were oxidatively modified in all body systems within the worm. Here, with the use of chemical penetration enhancers (CPEs), we increased the number of modified proteins as well as the number of modifications per protein to gain more structural information. CPEs aid in the delivery of hydrogen peroxide inside C. elegans by disturbing the highly ordered lipid bilayer of the worm cuticle without affecting worm viability. IV-FPOP experiments performed using the CPE azone showed an increase in oxidatively modified proteins and peptides. This increase correlated with greater hydrogen peroxide uptake by C. elegans quantified using a chemical fluorophore demonstrating the efficacy of using CPEs with IV-FPOP. Mass spectrometry proteomics data are available via ProteomeXchange with identifier PXD019290.

中文翻译：

化学渗透促进剂可增加线虫中过氧化氢的吸收，从而实现蛋白质的体内快速光化学氧化。

蛋白质的快速光化学氧化（FPOP）是一种羟基自由基蛋白质足迹方法，通过过氧化氢的光解作用共价标记溶剂可访问的氨基酸。最近，我们扩大了秀丽隐杆线虫体内（IV-FPOP）共价标记的FPOP用途。在最初的IV-FPOP研究中，蠕虫内所有身体系统中的545种蛋白质都被氧化修饰。在这里，通过使用化学渗透促进剂（CPE），我们增加了修饰蛋白的数量以及每种蛋白的修饰数量，以获得更多的结构信息。CPE有助于秀丽线虫内部过氧化氢的输送通过干扰蠕虫角质层的高度有序的脂质双层而不影响蠕虫的生存能力。使用CPE氮酮进行的IV-FPOP实验显示，氧化修饰的蛋白质和肽段增加。该增加与使用化学荧光团定量的秀丽隐杆线虫对更大的过氧化氢的吸收有关，这证明了将IV-FPOP与CPE一起使用的功效。质谱蛋白质组学数据可通过ProteomeXchange获得，其标识符为PXD019290。

更新日期：2020-06-07

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