Abstract
Metal ions are critical for the biological and physiological functions of many proteins. Mass spectrometry (MS)-based structural proteomics is an ever-growing field that has been adopted to study protein and metal ion interactions. Native MS offers information on metal binding and its stoichiometry. Footprinting approaches coupled with MS, including hydrogen/deuterium exchange (HDX), “fast photochemical oxidation of proteins” (FPOP) and targeted amino-acid labeling, identify binding sites and regions undergoing conformational changes. MS-based titration methods, including “protein–ligand interactions by mass spectrometry, titration and HD exchange” (PLIMSTEX) and “ligand titration, fast photochemical oxidation of proteins and mass spectrometry” (LITPOMS), afford binding stoichiometry, binding affinity, and binding order. These MS-based structural proteomics approaches, their applications to answer questions regarding metal ion protein interactions, their limitations, and recent and potential improvements are discussed here. This review serves as a demonstration of the capabilities of these tools and as an introduction to wider applications to solve other questions.
Highlights
Metal ions are critical for the biological and physiological functions of many proteins
After completion the+32 labeling, the reagent-responsive ensure that the footprinting occurs faster than protein folding or unfolding, the lifetim primary reactive hydroxyl radicals is limited to ~1 μs depending on the scavenger’s id tity and concentration [18,21], even though the lifetimes of less reactive secondary radi solvent-accessible residues contain a mass tag corresponding to the labeling product
Motivated by previous findings [30,31] indicating that divalent metals are necessary for the segmented negative-sense RNA virus’ endonuclease activity, the authors investigated the interaction between the endonuclease domain of severe fever with thrombocytopenia syndrome virus (SFTSV) L-polymerase (SFTSV endonuclease) and metal ions [28]
Summary
Metal ions play an essential role in biological and physiological processes, including respiration, signal transduction, and transcription [1,2]. MS-based structural proteomics tools bridge the gap between low- and high-resolution structural characterization and offer binding affinity and stoichiometry. We discuss the principles of MS-based structural proteomics tools; we illustrate their capability to characterize metal–protein interactions by discussing examples principally from our own work and some from other groups. These applications have been organized according to the different questions, in which biochemists and biophysicists might be interested, and the corresponding MS tools that can be used to answer these questions. Limitations of and principles for choosing each technique are discussed
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