Effect of DMSO on Protein Structure and Interactions Assessed by Collision-Induced Dissociation and Unfolding.
McLean, Kirsty J
American Chemical Society
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Chan, D., Kavanagh, M., McLean, K. J., Munro, A. W., Matak-Vinković, D., Coyne, A., & Abell, C. (2017). Effect of DMSO on Protein Structure and Interactions Assessed by Collision-Induced Dissociation and Unfolding.. Analytical chemistry, 89 (18), 9976-9983. https://doi.org/10.1021/acs.analchem.7b02329
Given the frequent use of DMSO in biochemical and biophysical assays, it is desirable to understand the influence of DMSO concentration on the dissociation or unfolding behavior of proteins. In this study, the effects of DMSO on the structure and interactions of avidin and Mycobacterium tuberculosis (Mtb) CYP142A1 were assessed through collision-induced dissociation (CID) and collision-induced unfolding (CIU) as monitored by nanoelectrospray ionization–ion mobility–mass spectrometry (nESI-IM-MS). DMSO concentrations higher than 4% (v/v) destabilize the avidin tetramer toward dissociation and unfolding, via both its effects on charge state distribution (CSD) as well as at the level of individual charge states. In contrast, DMSO both protects against heme loss and increases the stability of CYP142A1 toward unfolding even up to 40% DMSO. Tandem MS/MS experiments showed that DMSO could modify the dissociation pathway of CYP142A1, while CIU revealed the protective effect of the heme group on the structure of CYP142A1.
Mycobacterium tuberculosis, Dimethyl Sulfoxide, Cytochrome P-450 Enzyme System, Avidin, Spectrometry, Mass, Electrospray Ionization, Protein Conformation, Tandem Mass Spectrometry, Protein Unfolding
D.S.-H.C. acknowledges the Croucher Foundation and the Cambridge Commonwealth, European and International Trust for receipt of a Croucher Cambridge International Scholarship. M.E.K. was supported by a Commonwealth (University of Cambridge) Scholarship awarded in conjunc-tion with the Cambridge Commonwealth Trust and Cam-bridge Overseas Trust. K.J.M. and A.G.C. were supported by grants from the UK BBSRC (Biotechnology and Biological Sciences Research Council (BB/I019669/1 and BB/I019227/1).
External DOI: https://doi.org/10.1021/acs.analchem.7b02329
This record's URL: https://www.repository.cam.ac.uk/handle/1810/267874