title: Kinetics and mass spectrometric measurements of myoglobin unfolding in aqueous ionic liquid solutions.
abstract: Recent studies have characterized the effects of aqueous ionic liquids on myoglobin unfolding for the broader purposes of understanding their effects on protein structures, stabilities, and ultimately biocompatibilities for future applications. Here, we investigated the effects of four different ionic liquids (ILs) on the thermal stability, unfolding kinetics, and tertiary shape of myoglobin. We compared results for four different ILs: 1-butyl-3-methyl imidazolium tetrafluoroborate (BMIBF4); 1-butyl-3-methyl pyrrolidinium tetrafluoroborate (PyrrBF4); 1-ethyl-3-methyl imidazolium acetate (EMIAc); and tetramethylguanidinium acetate (TMGAc). Results showed that ILs accelerate myoglobin unfolding kinetics both through aqueous solution ionic strength effects and ionic liquid-specific effects. Arrhenius plots of observed rate constants reveal that some ILs lower the energy barrier to unfolding, possibly by destabilizing the native protein state. The magnitude of these ionic liquid effects correlates with their effects on protein thermodynamic stabilities. Hydrogen-deuterium exchange (HDX) experiments using ESI-MS showed that myoglobin exhibits a more open, and presumably less stable, tertiary shape in aqueous IL solutions. Overall, BMIBF4 and TMGAc exhibit the strongest effect on the myoglobin stability, unfolding rate, and tertiary structure while PyrrBF4 and EMIAc have weaker effects under our experimental conditions.
authors.affiliation: Department of Chemistry and Biochemistry, Rowan University, 201 Mullica Hill Road, Glassboro, NJ 08028, United States.
authors.forename: Miranda C
authors.initials: MC
authors.lastname: Miller
authors.affiliation: Department of Chemistry and Biochemistry, Rowan University, 201 Mullica Hill Road, Glassboro, NJ 08028, United States.
authors.forename: Sylvia L
authors.initials: SL
authors.lastname: Hanna
authors.affiliation: Department of Chemistry and Biochemistry, Rowan University, 201 Mullica Hill Road, Glassboro, NJ 08028, United States.
authors.forename: Kelsey G
authors.initials: KG
authors.lastname: DeFrates
authors.affiliation: Department of Chemistry and Biochemistry, Rowan University, 201 Mullica Hill Road, Glassboro, NJ 08028, United States.
authors.forename: Olivia C
authors.initials: OC
authors.lastname: Fiebig
authors.affiliation: Department of Chemistry and Biochemistry, Rowan University, 201 Mullica Hill Road, Glassboro, NJ 08028, United States. Electronic address: vadent@rowan.edu.
authors.forename: Timothy D
authors.initials: TD
authors.lastname: Vaden
mesh_terms: D000818:Animals; D006736:Horses; D006863:Hydrogen-Ion Concentration; D052578:Ionic Liquids / Q000737:chemistry*; D007700:Kinetics; D013058:Mass Spectrometry; D009211:Myoglobin / Q000737:chemistry*; D055550:Protein Stability; D058767:Protein Unfolding; D013816:Thermodynamics
chemical_list: D052578:Ionic Liquids; D009211:Myoglobin
keywords: Ionic liquids; Myoglobin; Protein unfolding