title: Fluorescence and circular dichroism spectroscopy of cytochrome c in alkylammonium formate ionic liquids.
abstract: The structural stability of cytochrome c has been studied in alkylammonium formate (AAF) ionic liquids such as methylammonium formate (MAF) and ethylammonium formate (EAF) by fluorescence and circular dichroism (CD) spectroscopy. At room temperature, the native structure of cytochrome c is maintained in relatively high ionic liquid concentrations (50-70% AAF/water or AAF/phosphate buffer pH 7.0) in contrast with denaturation of cytochrome c in similar solutions of methanol or acetonitrile with water or buffer cosolvents. Fluorescence and CD spectra indicate that the conformation of cytochrome c is maintained in 20% AAF-80% water from 30 to 50 °C. No such temperature stability is found in 80% AAF-20% water. About one-third of the enzyme activity of cytochrome c in 80% AAF-20% water can be maintained as compared with phosphate buffer, and this is greater than the activities measured in corresponding methanol and acetonitrile aqueous solutions. This biophysical study shows that AAFs have potential application as organic solvent replacements at moderate temperature in the mobile phase for the separation of proteins in their native form by reversed phase liquid chromatography.
authors.affiliation: Department of Chemistry and Biochemistry, Miami University, Oxford, Ohio 45056, USA.
authors.forename: Wenjun
authors.initials: W
authors.lastname: Wei
authors.forename: Neil D
authors.initials: ND
authors.lastname: Danielson
mesh_terms: D002942:Circular Dichroism; D045304:Cytochromes c / Q000737:chemistry*; D005561:Formates / Q000737:chemistry*; D052578:Ionic Liquids / Q000737:chemistry*; D008744:Methylamines / Q000737:chemistry*; D000644:Quaternary Ammonium Compounds / Q000737:chemistry*; D013050:Spectrometry, Fluorescence
chemical_list: D005561:Formates; D052578:Ionic Liquids; D008744:Methylamines; D000644:Quaternary Ammonium Compounds; C421484:ethylammonium; D045304:Cytochromes c; C027451:methylamine