Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents
The unique electrochemical properties of ionic liquids (ILs) have motivated their use as solvents for organic synthesis and green energy applications. More recently, their potential in pharmaceutical chemistry has prompted investigation into their effects on biomolecules. There is evidence that some...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-07-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/9/7/264 |
| _version_ | 1819037030124879872 |
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| author | Joshua Y. Lee Katherine M. Selfridge Eric M. Kohn Timothy D. Vaden Gregory A. Caputo |
| author_facet | Joshua Y. Lee Katherine M. Selfridge Eric M. Kohn Timothy D. Vaden Gregory A. Caputo |
| author_sort | Joshua Y. Lee |
| collection | DOAJ |
| description | The unique electrochemical properties of ionic liquids (ILs) have motivated their use as solvents for organic synthesis and green energy applications. More recently, their potential in pharmaceutical chemistry has prompted investigation into their effects on biomolecules. There is evidence that some ILs can destabilize proteins via a detergent-like manner; however, the mechanism still remains unknown. Our hypothesis is that if ILs are denaturing proteins via a detergent-like mechanism, detergent-mediated protein unfolding should be enhanced in the presence of ILs. The properties of myoglobin was examined in the presence of a zwitterionic (<i>N</i>,<i>N</i>-dimethyl-<i>N</i>-dodecylglycine betaine (Empigen BB<sup>®</sup>, EBB)), cationic (tetradecyltrimethylammonium bromide (TTAB)), and anionic (sodium dodecyl sulfate (SDS)) detergent as well as ILs based on alkylated imidazolium chlorides. Protein structure was measured through a combination of absorbance, fluorescence, and circular dichroism (CD) spectroscopy: absorbance and CD were used to monitor heme complexation to myoglobin, and tryptophan fluorescence quenching was used as an indicator for heme dissociation. Notably, the detergents tested did not fully denature the protein but instead resulted in loss of the heme group. At low IL concentrations, heme dissociation remained a traditional, cooperative process; at high concentrations, ILs with increased detergent-like character exhibited a more complex pattern, which is most likely attributable to micellization of the ionic liquids or direct denaturation or heme dissociation induced by the ILs. These trends were consistent across all species of detergents. 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence was further used to characterize micelle formation in aqueous solutions containing detergent and ionic liquid. The dissociation thermodynamics show that EBB- and TTAB-induced dissociation of heme is not significantly impacted by room temperature ionic liquids (RTILs), whereas SDS-induced dissociation is more dramatically impacted by all RTILs examined. Together, these results indicate a complex interaction of detergents, likely based on headgroup charge, and the active component of RTILs to influence heme dissociation and potentially protein denaturation. |
| first_indexed | 2024-12-21T08:14:56Z |
| format | Article |
| id | doaj.art-a54bf11961354eefb773c60c07de052d |
| institution | Directory Open Access Journal |
| issn | 2218-273X |
| language | English |
| last_indexed | 2024-12-21T08:14:56Z |
| publishDate | 2019-07-01 |
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| record_format | Article |
| series | Biomolecules |
| spelling | doaj.art-a54bf11961354eefb773c60c07de052d2022-12-21T19:10:35ZengMDPI AGBiomolecules2218-273X2019-07-019726410.3390/biom9070264biom9070264Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic DetergentsJoshua Y. Lee0Katherine M. Selfridge1Eric M. Kohn2Timothy D. Vaden3Gregory A. Caputo4Department of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USADepartment of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USADepartment of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USADepartment of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USADepartment of Chemistry and Biochemistry, Rowan University, Glassboro, NJ 08028, USAThe unique electrochemical properties of ionic liquids (ILs) have motivated their use as solvents for organic synthesis and green energy applications. More recently, their potential in pharmaceutical chemistry has prompted investigation into their effects on biomolecules. There is evidence that some ILs can destabilize proteins via a detergent-like manner; however, the mechanism still remains unknown. Our hypothesis is that if ILs are denaturing proteins via a detergent-like mechanism, detergent-mediated protein unfolding should be enhanced in the presence of ILs. The properties of myoglobin was examined in the presence of a zwitterionic (<i>N</i>,<i>N</i>-dimethyl-<i>N</i>-dodecylglycine betaine (Empigen BB<sup>®</sup>, EBB)), cationic (tetradecyltrimethylammonium bromide (TTAB)), and anionic (sodium dodecyl sulfate (SDS)) detergent as well as ILs based on alkylated imidazolium chlorides. Protein structure was measured through a combination of absorbance, fluorescence, and circular dichroism (CD) spectroscopy: absorbance and CD were used to monitor heme complexation to myoglobin, and tryptophan fluorescence quenching was used as an indicator for heme dissociation. Notably, the detergents tested did not fully denature the protein but instead resulted in loss of the heme group. At low IL concentrations, heme dissociation remained a traditional, cooperative process; at high concentrations, ILs with increased detergent-like character exhibited a more complex pattern, which is most likely attributable to micellization of the ionic liquids or direct denaturation or heme dissociation induced by the ILs. These trends were consistent across all species of detergents. 1,6-diphenyl-1,3,5-hexatriene (DPH) fluorescence was further used to characterize micelle formation in aqueous solutions containing detergent and ionic liquid. The dissociation thermodynamics show that EBB- and TTAB-induced dissociation of heme is not significantly impacted by room temperature ionic liquids (RTILs), whereas SDS-induced dissociation is more dramatically impacted by all RTILs examined. Together, these results indicate a complex interaction of detergents, likely based on headgroup charge, and the active component of RTILs to influence heme dissociation and potentially protein denaturation.https://www.mdpi.com/2218-273X/9/7/264room temperature ionic liquidsfluorescenceprotein foldingionic liquidshememyoglobin |
| spellingShingle | Joshua Y. Lee Katherine M. Selfridge Eric M. Kohn Timothy D. Vaden Gregory A. Caputo Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents Biomolecules room temperature ionic liquids fluorescence protein folding ionic liquids heme myoglobin |
| title | Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents |
| title_full | Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents |
| title_fullStr | Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents |
| title_full_unstemmed | Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents |
| title_short | Effects of Ionic Liquid Alkyl Chain Length on Denaturation of Myoglobin by Anionic, Cationic, and Zwitterionic Detergents |
| title_sort | effects of ionic liquid alkyl chain length on denaturation of myoglobin by anionic cationic and zwitterionic detergents |
| topic | room temperature ionic liquids fluorescence protein folding ionic liquids heme myoglobin |
| url | https://www.mdpi.com/2218-273X/9/7/264 |
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