Abstract
Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS) is a rapidly evolving technique for analyzing structural features and dynamic properties of proteins. It may stand alone or serve as a complementary method to cryo-electron-microscopy (EM) or other structural biology approaches. HDX-MS is capable of providing information on individual proteins as well as large protein complexes. Owing to recent methodological advancements and improving availability of instrumentation, HDX-MS is becoming a routine technique for some applications. When dealing with samples of low to medium complexity and sizes of less than 150 kDa, conformation and ligand interaction analyses by HDX-MS are already almost routine applications. This is also well supported by the rapid evolution of the computational (software) background that facilitates the analysis of the obtained experimental data. HDX-MS can cope at times with analytes that are difficult to tackle by any other approach. Large complexes like viral capsids as well as disordered proteins can also be analyzed by this method. HDX-MS has recently become an established tool in the drug discovery process and biopharmaceutical development, as it is now also capable of dissecting post-translational modifications and membrane proteins. This mini review provides the reader with an introduction to the technique and a brief overview of the most common applications. Furthermore, the most challenging likely applications, the analyses of glycosylated and membrane proteins, are also highlighted.
Highlights
The origins of hydrogen-deuterium exchange date back to the middle of the last century [1,2], when the technique was first applied in the field of NMR spectroscopy to study molecular dynamics.Mass spectrometry came later to this field, in the 1990s, when it became feasible to study large proteins and their complexes
The degree of accord of the results show how reliable the Hydrogen/Deuterium eXchange Mass Spectrometry (HDX-MS) results are for a protein
This study demonstrated temperature-dependent dynamic hotspots on the surface of the capsid and concluded that HDX-MS had successfully been applied to determine potential epitopes on the virus
Summary
The origins of hydrogen-deuterium exchange date back to the middle of the last century [1,2], when the technique was first applied in the field of NMR spectroscopy to study molecular dynamics. Hydrogen-Deuterium eXchange Mass Spectrometry (HDX-MS) is a powerful technique for structural protein science, providing detailed insights into protein structure as well as conformational dynamics and function [3] This approach has already become a frequently applied and well-established methodology in the field of protein structure. Unlike several other biophysical techniques, HDX-MS possesses the advantages of no or very high size limit [8] and is useful for studying individual proteins as well as large complexes [34,35] This is mainly due to the application of proteolysis [14] and radical-induced fragmentation (electron transfer and electron capture dissociation respectively; ETD and ECD) in the course of analysis [36,37,38]. This review is a short overview of the HDX-MS method, which focuses on the strengths and weaknesses of the technique as well as data interpretation
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