In-Cell Fast Photochemical Oxidation of Integral Membrane Proteins

We introduce IC-FPOMP (in-cell fast photochemical oxidation of MPs), a method enabling in situ footprinting of IMPs within live cells. IC-FPOMP generates reactive oxygen radicals from several precursors nearby the membrane. We achieve high-throughput and rapid footprinting of IMPs. IC-FPOMP of two human IMPs (human glucose transporter-hGLUT1 and human gamma glutamyl carboxylase-hGGCX) were successful, providing comprehensive footprinting of both the transmembrane and extramembrane regions. IC-FPOMP offers insights on IMP structure-function relationships in cell for the first time, facilitating drug discovery.

本研究介绍了IC-FPOMP（胞内快速光化学氧化膜蛋白法，in-cell fast photochemical oxidation of MPs），这是一种可实现活细胞内整合膜蛋白（integral membrane proteins，IMPs）原位足迹分析的技术方法。该方法可通过膜附近的多种前驱体产生活性氧自由基，能够实现整合膜蛋白的高通量、快速足迹分析。针对两种人类整合膜蛋白——人葡萄糖转运蛋白hGLUT1（human glucose transporter-hGLUT1）与人γ-谷氨酰羧化酶hGGCX（human gamma glutamyl carboxylase-hGGCX）的IC-FPOMP实验均取得成功，实现了这两种蛋白跨膜区与膜外区的全面足迹分析。该方法首次为解析活细胞内整合膜蛋白的结构-功能关联提供了关键研究视角，有助于推动药物研发进程。