Open AccessFast Photochemical Oxidation of Proteins Coupled with Mass Spectrometry
Author(s) -
Liuqing Shi,
Michael L. Gross
Publication year - 2019
Publication title -
protein and peptide letters/protein and peptide letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.432
H-Index - 58
eISSN - 1875-5305
pISSN - 0929-8665
DOI - 10.2174/0929866526666181128124554
Subject(s) - chemistry , footprinting , radical , mass spectrometry , macromolecule , hydroxyl radical , dna footprinting , proteomics , protein structure , quantitative proteomics , combinatorial chemistry , biochemistry , chromatography , dna , dna binding protein , transcription factor , base sequence , gene
Determination of the composition and some structural features of macromolecules can be achieved by using structural proteomics approaches coupled with mass spectrometry (MS). One approach is hydroxyl radical protein footprinting whereby amino-acid side chains are modified with reactive reagents to modify irreversibly a protein side chain. The outcomes, when deciphered with mass-spectrometry-based proteomics, can increase our knowledge of structure, assembly, and conformational dynamics of macromolecules in solution. Generating the hydroxyl radicals by laser irradiation, Hambly and Gross developed the approach of Fast Photochemical Oxidation of Proteins (FPOP), which labels proteins on the sub millisecond time scale and provides, with MS analysis, deeper understanding of protein structure and protein-ligand and protein- protein interactions. This review highlights the fundamentals of FPOP and provides descriptions of hydroxyl-radical and other radical and carbene generation, of the hydroxyl labeling of proteins, and of determination of protein modification sites. We also summarize some recent applications of FPOP coupled with MS in protein footprinting.