A Mass Spectrometry Study to Monitor the Conformation Change of Protein in Deep Eutectic Solvent
Disciplines
Analytical Chemistry
Abstract (300 words maximum)
Deep eutectic solvents (DESs), an emerging class of green solvents, are formed by mixing two components at a certain composition. DESs are considered biodegradable, sustainable, and non-toxic in nature. Most protein and enzyme solubility and reaction studies are performed in aqueous or organic solvents. However, DESs can affect the structure and dynamics of the protein and enzyme have yet to be explored. The aim of this study to explore the DES-induced protein and enzyme structural changes using mass spectrometry. In this experiment, Lysozyme and α-Lactalbumin are used as model proteins. Two different DESs including menthol: acetic acid and proline: urea were prepared for this experiment. The formation of DESs is confirmed by infrared spectroscopy. Initially Lysozyme sample was prepared at 10 μM concentration using LC-MS grade water. In addition, separately, protein samples were prepared with 1% -30% menthol: acetic acid and proline:urea DESs. Lysozyme in water showed the charge state distribution from 6+ to 13+ , in which the 9+ being the most intense peak. When 1% acetic acid was added to the Lysozyme solution, charged states distribution changed from 5+ to 13+, where the most intense peak noticed for 9+ . This indicates that the Lysozyme slightly folded by the interactions of acetic acid molecules compared to its native state. However, in 1% menthol: acetic acid DES solution, Lysozyme was more folded compared to water. In addition, the ion current of Lysozyme charge states is more noticeable in Men:AA compared to water. At 30% of menthol: acetic acid DES, the Lysozyme was slightly unfolded as the charge states distribution was shifted. However, the impact of amino acid based proline:urea DES on protein structure and dynamics is very different in which the charge stated distribution of α-Lactalbumin was shifted to low m/z and adopted more folded states compared to water.
Academic department under which the project should be listed
CSM - Chemistry and Biochemistry
Primary Investigator (PI) Name
Mohammad A. Halim
A Mass Spectrometry Study to Monitor the Conformation Change of Protein in Deep Eutectic Solvent
Deep eutectic solvents (DESs), an emerging class of green solvents, are formed by mixing two components at a certain composition. DESs are considered biodegradable, sustainable, and non-toxic in nature. Most protein and enzyme solubility and reaction studies are performed in aqueous or organic solvents. However, DESs can affect the structure and dynamics of the protein and enzyme have yet to be explored. The aim of this study to explore the DES-induced protein and enzyme structural changes using mass spectrometry. In this experiment, Lysozyme and α-Lactalbumin are used as model proteins. Two different DESs including menthol: acetic acid and proline: urea were prepared for this experiment. The formation of DESs is confirmed by infrared spectroscopy. Initially Lysozyme sample was prepared at 10 μM concentration using LC-MS grade water. In addition, separately, protein samples were prepared with 1% -30% menthol: acetic acid and proline:urea DESs. Lysozyme in water showed the charge state distribution from 6+ to 13+ , in which the 9+ being the most intense peak. When 1% acetic acid was added to the Lysozyme solution, charged states distribution changed from 5+ to 13+, where the most intense peak noticed for 9+ . This indicates that the Lysozyme slightly folded by the interactions of acetic acid molecules compared to its native state. However, in 1% menthol: acetic acid DES solution, Lysozyme was more folded compared to water. In addition, the ion current of Lysozyme charge states is more noticeable in Men:AA compared to water. At 30% of menthol: acetic acid DES, the Lysozyme was slightly unfolded as the charge states distribution was shifted. However, the impact of amino acid based proline:urea DES on protein structure and dynamics is very different in which the charge stated distribution of α-Lactalbumin was shifted to low m/z and adopted more folded states compared to water.