Abstract
Electrospray ionization mass spectrometry (ESI-MS) is nowadays one of the cornerstones of biomolecular mass spectrometry and proteomics. Advances in sample preparation and mass analyzers have enabled researchers to extract much more information from biological samples than just the molecular weight. In particular, relevant for structural biology, noncovalent protein–protein and protein–ligand complexes can now also be analyzed by MS. For these types of analyses, assemblies need to be retained in their native quaternary state in the gas phase. This initial small niche of biomolecular mass spectrometry, nowadays often referred to as “native MS,” has come to maturation over the last two decades, with dozens of laboratories using it to study mostly protein assemblies, but also DNA and RNA-protein assemblies, with the goal to define structure–function relationships. In this perspective, we describe the origins of and (re)define the term native MS, portraying in detail what we meant by “native MS,” when the term was coined and also describing what it does (according to us) not entail. Additionally, we describe a few examples highlighting what native MS is, showing its successes to date while illustrating the wide scope this technology has in solving complex biological questions.Graphical ᅟ
Highlights
IntroductionDifferent methods of mass spectrometry (compared with Mass spectrometry (MS)-based proteomics analysis) are rapidly emerging that are complimentary to the aforementioned biophysical techniques for structural biology, enabling investigations into tertiary and quaternary structures of protein assemblies
All cellular biological processes involve the complex coordination of proteins and other biomolecules (e.g., DNA, RNA, lipids, metabolites) in space and time
Mass spectrometry (MS), MS-based proteomics, has already played a pivotal role in deciphering the many protein complexes and protein interaction networks that are present in the cell, typically at the end of a pipe-line that starts with the immune-purification of a tagged-bait protein, followed by identification of the co-purified proteins using LC-MS/MS [5, 6]
Summary
Different methods of mass spectrometry (compared with MS-based proteomics analysis) are rapidly emerging that are complimentary to the aforementioned biophysical techniques for structural biology, enabling investigations into tertiary and quaternary structures of protein assemblies. These span the range from hydrogen/deuterium exchange mass spectrometry, chemical surface labeling techniques, cross-linking mass spectrometry, up to the direct mass analysis of intact protein assemblies [9] The latter arose following initial work by a few laboratories in the 1990s [10, 11], demonstrating that noncovalent interactions could be preserved in the gas phase for analysis, enabling information on subunit stoichiometry, binding partners, protein complex topology, protein dynamics, and even binding affinities from a single mass spectrometric analysis [12,13,14,15,16,17]. Conference, BNative Mass Spectrometry-Based Structural Biology^ held in the autumn of 2015
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
