Abstract
Ion charge-state distribution analysis in electro-spray ionization mass spectrometry (ESI-MS) is a robust and fast technique for direct detection and characterization of coexisting protein conformations in solution. Compact folded proteins give rise to ESI-generated ions carrying a relatively small number of charges, whereas less compact conformers accommodate upon ESI a larger number of charges depending on the extent of their unfolding. A chemometric approach [1] based upon factor analysis is applied to determine contributions from individual conformers to the overall CSD. Here we present basic guidelines for the use of this MS-based technique: from the preparation of suitable solutions for ESI-MS to the acquisition of reliable MS data and their subsequent analysis.
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References
Mohimen A, Dobo A, Hoerner JK, Kaltashov IA (2003) A chemometric approach to detection and characterization of multiple protein conformers in solution using electrospray ionization mass spectrometry. Anal Chem 75:4139–4147
Konermann L, Douglas DJ (1998) Equilibrium unfolding of proteins monitored by electrospray ionization mass spectrometry: distinguishing two-state from multi-state transitions. Rapid Commun Mass Spectrom 12:435–442
Frimpong AK, Abzalimov RR, Eyles SJ, Kaltashov IA (2007) Gas-phase interference-free analysis of protein ion charge-state distributions: detection of small-scale conformational transitions accompanying pepsin inactivation. Anal Chem 79:4154–4161
Invernizzi G, Grandori R (2007) Detection of the equilibrium folding intermediate of beta-lactoglobulin in the presence of trifluoroethanol by mass spectrometry. Rapid Commun Mass Spectrom 21:1049–1052
Zhang M, Gumerov DR, Kaltashov IA, Mason AB (2004) Indirect detection of protein-metal binding: interaction of serum transferrin with In3+ and Bi3+. J Am Soc Mass Spectrom 15:1658–1664
Griffith WP, Kaltashov IA (2003) Highly asymmetric interactions between globin chains during hemoglobin assembly revealed by electrospray ionization mass spectrometry. Biochemistry 42:10024–10033
Simmons DA, Wilson DJ, Lajoie GA, Doherty-Kirby A, Konermann L (2004) Subunit disassembly and unfolding kinetics of hemoglobin studied by time-resolved electrospray mass spectrometry. Biochemistry 43:14792–14801
Gumerov DR, Dobo A, Kaltashov IA (2002) Protein-ion charge-state distributions in electrospray ionization mass spectrometry: distinguishing conformational contributions from masking effects. Eur J Mass Spectrom 8:123–129
Dobo A, Kaltashov IA (2001) Detection of multiple protein conformational ensembles in solution via deconvolution of charge state distributions in ESI MS. Anal Chem 73:4763–4773
Frimpong AK, Abzalimov RR, Uversky VN, Kaltashov IA (2010) Characterization of intrinsically disordered proteins with electrospray ionization mass spectrometry: conformational heterogeneity of α-synuclein. Proteins 78(3):714–722
Watkins DS (2005) Fundamentals of matrix computations, 2nd edn. http://onlinelibrary.wiley.com/book/10.1002/0471249718
Kaltashov IA, Eyles SJ, Mohimen A, Hoerner JK, Abzalimov RR, Griffith WP (2007) NMR and EPR Spectroscopies, Mass-Spectrometry and Protein Imaging, In: Uversky V, Eugene A. Permyakov (eds), Methods in Protein Structure and Stability Analysis. Nova science, U S A. ISBN 978-1-60021-705-0, 175–196
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This work was supported by a grant CHE-0750389 from the National Science Foundation.
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Abzalimov, R.R., Frimpong, A.K., Kaltashov, I.A. (2012). Detection and Characterization of Large-Scale Protein Conformational Transitions in Solution Using Charge-State Distribution Analysis in ESI-MS. In: Uversky, V., Dunker, A. (eds) Intrinsically Disordered Protein Analysis. Methods in Molecular Biology, vol 896. Springer, New York, NY. https://doi.org/10.1007/978-1-4614-3704-8_24
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DOI: https://doi.org/10.1007/978-1-4614-3704-8_24
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