title: Surfactant effects on protein structure examined by electrospray ionization mass spectrometry.
abstract: Electrospray ionization mass spectrometry (ESI-MS) has proven to be a useful tool for examining noncovalent complexes between proteins and a variety of ligands. It has also been used to distinguish between denatured and refolded forms of proteins. Surfactants are frequently employed to enhance solubilization or to modify the tertiary or quaternary structure of proteins, but are usually considered incompatible with mass spectrometry. A broad range of ionic, nonionic, and zwitterionic surfactants was examined to characterize their effects on ESI-MS and on protein structure under ESI-MS conditions. Solution conditions studied include 4% acetic acid/50% acetonitrile/46% H2O and 100% aqueous. Of the surfactants examined, the nonionic saccharides, such as n-dodecyl-beta-D-glucopyranoside, at 0.1% to 0.01% (w/v) concentrations, performed best, with limited interference from chemical background and adduct formation. Under the experimental conditions used, ESI-MS performance in the presence of surfactants was found to be unrelated to critical micelle concentration. It is demonstrated that surfactants can affect both the tertiary and quaternary structures of proteins under conditions used for ESI-MS. However, several of the surfactants caused significant shifts in the charge-state distributions, which appeared to be independent of conformational effects. These observations suggest that surfactants, used in conjunction with ESI-MS, can be useful for protein structure studies, if care is used in the interpretation of the results.
authors.affiliation: Department of Biological Chemistry, University of Michigan, Ann Arbor 48109.
authors.forename: R R
authors.initials: RR
authors.lastname: Loo
authors.forename: N
authors.initials: N
authors.lastname: Dales
authors.forename: P C
authors.initials: PC
authors.lastname: Andrews
mesh_terms: D003574:Cytochrome c Group / Q000737:chemistry; D003902:Detergents / Q000494:pharmacology; D013058:Mass Spectrometry / Q000379:methods; D008823:Micelles; D009113:Muramidase / Q000737:chemistry; D009211:Myoglobin / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation / Q000187:drug effects*; D011489:Protein Denaturation; D017434:Protein Structure, Tertiary / Q000187:drug effects*; D013501:Surface-Active Agents / Q000494:pharmacology*; D014452:Ubiquitins / Q000737:chemistry*
chemical_list: D003574:Cytochrome c Group; D003902:Detergents; D008823:Micelles; D009211:Myoglobin; D013501:Surface-Active Agents; D014452:Ubiquitins; D009113:Muramidase