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.
Original language | English (US) |
---|---|
Pages (from-to) | 1061-1068 |
Number of pages | 8 |
Journal | Protein Science |
Volume | 25 |
Issue number | 5 |
DOIs | |
State | Published - May 1 2016 |
Keywords
- circular dichroism
- dodine acetate
- guanidine hydrochloride
- infrared spectroscopy
- phosphoglycerate kinase
- pressure denaturation
- protein folding
- thermal denaturation
- tryptophan fluorescence
ASJC Scopus subject areas
- Biochemistry
- Molecular Biology