title: Ion mobility-mass spectrometry reveals conformational changes in charge reduced multiprotein complexes.
abstract: Characterizing intact multiprotein complexes in terms of both their mass and size by ion mobility-mass spectrometry is becoming an increasingly important tool for structural biology. Furthermore, the charge states of intact protein complexes can dramatically influence the information content of gas-phase measurements performed. Specifically, protein complex charge state has a demonstrated influence upon the conformation, mass resolution, ion mobility resolution, and dissociation properties of protein assemblies upon collisional activation. Here we present the first comparison of charge-reduced multiprotein complexes generated by solution additives and gas-phase ion-neutral reaction chemistry. While the charge reduction mechanism for both methods is undoubtedly similar, significant gas-phase activation of the complex is required to reduce the charge of the assemblies generated using the solution additive strategy employed here. This activation step can act to unfold intact protein complexes, making the data difficult to correlate with solution-phase structures and topologies. We use ion mobility-mass spectrometry to chart such conformational effects for a range of multi-protein complexes, and demonstrate that approaches to reduce charge based on ion-neutral reaction chemistry in the gas-phase consistently produce protein assemblies having compact, 'native-like' geometries while the same molecules added in solution generate significantly unfolded gas-phase complexes having identical charge states.
authors.affiliation: Department of Chemistry, University of Michigan, 930 N. University Ave., Ann Arbor, MI 48109, USA.
authors.forename: Russell E
authors.initials: RE
authors.lastname: Bornschein
authors.forename: Suk-Joon
authors.initials: SJ
authors.lastname: Hyung
authors.forename: Brandon T
authors.initials: BT
authors.lastname: Ruotolo
mesh_terms: D000818:Animals; D002645:Chickens; D005740:Gases; D007477:Ions; D046912:Multiprotein Complexes / Q000737:chemistry*; D036103:Nanotechnology; D011487:Protein Conformation; D011817:Rabbits; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
chemical_list: D005740:Gases; D007477:Ions; D046912:Multiprotein Complexes