Approaches to Heterogeneity in Native Mass Spectrometry

Native mass spectrometry (MS) is aimed at preserving and determining the native structure, composition, and stoichiometry of biomolecules and their complexes from solution after they are transferred into the gas phase. Major improvements in native MS instrumentation and experimental methods over the past few decades have led to a concomitant increase in the complexity and heterogeneity of samples that can be analyzed, including protein-ligand complexes, protein complexes with multiple coexisting stoichiometries, and membrane protein-lipid assemblies. Heterogeneous features of these biomolecular samples can be important for understanding structure and function. However, sample heterogeneity can make assignment of ion mass, charge, composition, and structure very challenging due to the overlap of tens or even hundreds of peaks in the mass spectrum. In this review, we cover data analysis, experimental, and instrumental advances and strategies aimed at solving this problem, with an in-depth discussion of theoretical and practical aspects of the use of available deconvolution algorithms and tools. We also reflect upon current challenges and provide a view of the future of this exciting field.

Automated summary

Native mass spectrometry is a method to preserve and determine the native structure, composition, and stoichiometry of biomolecules and their complexes after they are transferred into the gas phase. Recent advancements in instrumentation and experimental methods have allowed for the analysis of more complex and heterogeneous samples, providing insights into the structure and function of these biomolecules. However, the heterogeneity of the samples poses challenges in assigning ion mass, charge, composition, and structure, which can be addressed using deconvolution algorithms and tools.