title: Applying mass spectrometry to study non-covalent biomolecule complexes.
abstract: Non-covalent interactions are essential for the structural organization of biomacromolecules and play an important role in molecular recognition processes, such as the interactions between proteins, glycans, lipids, DNA, and RNA. Mass spectrometry (MS) is a powerful tool for studying of non-covalent interactions, due to the low sample consumption, high sensitivity, and label-free nature. Nowadays, native-ESI MS is heavily used in studies of non-covalent interactions and to understand the architecture of biomolecular complexes. However, MALDI-MS is also becoming increasingly useful. It is challenging to detect the intact complex without fragmentation when analyzing non-covalent interactions with MALDI-MS. There are two methodological approaches to do so. In the first approach, different experimental and instrumental parameters are fine-tuned in order to find conditions under which the complex is stable, such as applying non-acidic matrices and collecting first-shot spectra. In the second approach, the interacting species are "artificially" stabilized by chemical crosslinking. Both approaches are capable of studying non-covalently bound biomolecules even in quite challenging systems, such as membrane protein complexes. Herein, we review and compare native-ESI and MALDI MS for the study of non-covalent interactions.
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
authors.forename: Fan
authors.initials: F
authors.lastname: Chen
authors.affiliation: Institute of Child Health, Division of Pediatric Basic Sciences, Hacettepe University, 06100 Ankara, Turkey.
authors.forename: Basri
authors.initials: B
authors.lastname: Gülbakan
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
authors.forename: Simon
authors.initials: S
authors.lastname: Weidmann
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
authors.forename: Stephan R
authors.initials: SR
authors.lastname: Fagerer
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
authors.forename: Alfredo J
authors.initials: AJ
authors.lastname: Ibáñez
authors.affiliation: Department of Chemistry and Applied Biosciences, ETH Zürich, CH-8093, Zürich, Switzerland.
authors.forename: Renato
authors.initials: R
authors.lastname: Zenobi
mesh_terms: D001671:Biochemistry / Q000379:methods*; D001703:Biophysics / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D018360:Crystallography, X-Ray / Q000379:methods; D005976:Glutaral / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D046912:Multiprotein Complexes / Q000032:analysis / Q000737:chemistry*; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods
chemical_list: D003432:Cross-Linking Reagents; D046912:Multiprotein Complexes; D005976:Glutaral
keywords: ESI; MALDI; chemical cross-linking; first shot phenomenon; non-covalent interactions