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Updated: Feb 10, 2026

Characterizing Cellular Proteins with In-cell Fast Photochemical Oxidation of Proteins
Published on: March 11, 2020
Implementing fast photochemical oxidation of proteins (FPOP) as a footprinting approach to solve diverse problems in
Bojie Zhang1, Ming Cheng1, Don Rempel1
1Department of Chemistry, Washington University in St. Louis, St. Louis, MO 63130, USA.
Abstract:
Fast photochemical oxidation of proteins (FPOP) is a footprinting technique used in mass spectrometry-based structural proteomics. It has been applied to solve a variety of problems in different areas of biology. A FPOP platform requires a laser, optics, and sample flow path properly assembled to enable fast footprinting. Sample preparation, buffer conditions, and reagent concentrations are essential to obtain reasonable oxidations on proteins. FPOP samples can be analyzed by LC-MS methods to measure the modification extent, which is a function of the solvent-accessible surface area of the protein. The platform can be expanded to accommodate several new approaches, including dose-response studies, new footprinting reagents, and two-laser pump-probe experiments. Here, we briefly review FPOP applications and in a detailed manner describe the procedures to set up an FPOP protein footprinting platform.
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