title: Dodine as a transparent protein denaturant for circular dichroism and infrared studies.
abstract: The fungicide dodine combines the cooperative denaturation properties of guanidine with the mM denaturation activity of SDS. It was previously tested only on two small model proteins. Here we show that it can be used as a chemical denaturant for phosphoglycerate kinase (PGK), a much larger two-domain enzyme. In addition to its properties as a chemical denaturant, dodine facilitates thermal denaturation of PGK, and we show for the first time that it also facilitates pressure denaturation of a protein. Much higher quality circular dichroism and amide I' infrared spectra of PGK can be obtained in dodine than in guanidine, opening the possibility for use of dodine as a denaturant when UV or IR detection is desirable. One caution is that dodine denaturation, like other detergent-based denaturants, is less reversible than guanidine denaturation.
authors.affiliation: Department of Chemistry, University of Illinois, Urbana, Illinois, 61801.
authors.forename: Drishti
authors.initials: D
authors.lastname: Guin
authors.affiliation: Department of Chemistry, North Central College, Naperville, Illinois, 60540.
authors.forename: Kori
authors.initials: K
authors.lastname: Sye
authors.affiliation: Center for Biophysics and Quantitative Biology, University of Illinois, Urbana, Illinois, 61801.
authors.forename: Kapil
authors.initials: K
authors.lastname: Dave
authors.affiliation: Department of Chemistry, University of Illinois, Urbana, Illinois, 61801.|Center for Biophysics and Quantitative Biology, University of Illinois, Urbana, Illinois, 61801.|Department of Physics, University of Illinois, Urbana, Illinois, 61801.
authors.forename: Martin
authors.initials: M
authors.lastname: Gruebele
mesh_terms: D002942:Circular Dichroism / Q000379:methods*; D019791:Guanidine / Q000737:chemistry; D006146:Guanidines / Q000737:chemistry*; D008958:Models, Molecular; D010735:Phosphoglycerate Kinase / Q000737:chemistry*; D011487:Protein Conformation; D011489:Protein Denaturation; D013055:Spectrophotometry, Infrared
chemical_list: D006146:Guanidines; C006921:dodine; D010735:Phosphoglycerate Kinase; D019791:Guanidine
keywords: circular dichroism; dodine acetate; guanidine hydrochloride; infrared spectroscopy; phosphoglycerate kinase; pressure denaturation; protein folding; thermal denaturation; tryptophan fluorescence