Abstract
Ion mobility spectrometry (IMS) coupled with mass spectrometry (MS) enables the investigation of protein folding in solution. Herein, a proof‐of‐concept for obtaining structural information about the folding of a protein in dependency of the amount of an organic cosolvent in the aqueous medium by means of this IMS‐MS method is presented. By analyzing the protein with native nano‐electrospray ionization IMS‐MS, the impact of acetonitrile as a representative organic cosolvent and/or pH values on the folding of an enzyme was successfully evaluated in a fast and straightforward fashion, as exemplified for an ene reductase from Gluconobacter oxydans. The IMS‐MS results are in agreement with findings from the nicotinamide adenine dinucleotide phosphate (NADPH)‐based spectrophotometric enzyme activity tests under analogous conditions, and thus, also rationalizing these “wet” analytical data. For this ene reductase, a higher tolerance against CH3CN in the presence of a buffer was observed by both analytical methods. The results suggest that this IMS‐MS methodology could be a useful complementary tool to existing methods in process optimization and fine‐tuning of solvent conditions for biotransformations.
Highlights
Ion mobility spectrometry (IMS) coupled with mass spectrometry (MS) enables the investigation of protein folding in solution
Encouraged by the success of ion mobility mass spectrometry (IMS) coupled with mass spectrometry (MS) for studying protein structures in the absence of bulk water,[3,4] we became interested in studying IMS-MS methodology[5,6] for obtaining information about the folding of this protein in dependency of the amount of an organic cosolvent in the aqueous medium
We report our results on such a comparison of the structural information about the enzyme obtained from IMS with those of the “wet” analytical data obtained from the standard spectrophotometric measurement of the enzyme activity under the same experimental conditions
Summary
Ion mobility spectrometry (IMS) coupled with mass spectrometry (MS) enables the investigation of protein folding in solution. In our study, using IMS, we were interested in gaining an insight into whether the decrease in activity was related to changes in enzyme folding caused by CH3CN or due to the pH of the aqueous solution.
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