Evaluation of capillary electrophoresis-mass spectrometry for the analysis of the conformational heterogeneity of intact proteins using beta2-microglobulin as model compound
| Autor: | Elena Domínguez-Vega, Julie Schappler, Laura Bertoletti, Raffaella Colombo, Serge Rudaz, Ersilia De Lorenzi, Sara Raimondi, Govert W. Somsen, Rob Haselberg |
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| Přispěvatelé: | BioAnalytical Chemistry, AIMMS |
| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
0301 basic medicine
Electrospray Chromatography Chemistry 010401 analytical chemistry Analytical chemistry Electrolyte Mass spectrometry 01 natural sciences Biochemistry Capillary electrophoresis–mass spectrometry 0104 chemical sciences Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound 030104 developmental biology Protein structure Ammonium bicarbonate Environmental Chemistry Conformational isomerism Spectroscopy Dynamic equilibrium |
| Zdroj: | Bertoletti, L, Schappler, J, Colombo, R, Rudaz, S, Haselberg, R, Dominguez Vega, E, Raimondi, S, Somsen, G W & De Lorenzi, E 2016, ' Evaluation of capillary electrophoresis-mass spectrometry for the analysis of the conformational heterogeneity of intact proteins using beta2-microglobulin as model compound. ', Analytica Chimica Acta, vol. 945, pp. 102-109 . https://doi.org/10.1016/j.aca.2016.10.010 Analytica Chimica Acta, 945, 102-109. Elsevier |
| ISSN: | 0003-2670 |
| Popis: | In this work we explored the feasibility of different CE-ESI-MS set-ups for the analysis of conformational states of an intact protein. By using the same background electrolyte at quasi physiological conditions (50 mM ammonium bicarbonate, pH 7.4) a sequential optimization was carried out, initially by evaluating a sheath-liquid interface with both a single quadrupole (SQ) and a time-of-flight (TOF) mass spectrometer; then a sheathless interface coupled with high-resolution QTOF MS was considered. Beta2-microglobulin has been taken as a model, as it is an amyloidogenic protein and its conformational changes are strictly connected to the onset of a disease. The separation of two conformers at dynamic equilibrium is achieved all the way down to the MS detection. Notably, the equilibrium ratio of the protein conformers is maintained in the electrospray source after CE separation. Strengths and weaknesses of each optimized set-up are emphasized and their feasibility in unfolding studies is evaluated. In particular, ESI-TOF MS can assign protein forms that differ by 1 Da only and sheathless interfacing is best suited to preserve protein structure integrity. This demonstrates the CE-ESI-MS performance in terms of separation, detection and characterization of conformational species that co-populate a protein solution. |
| Databáze: | OpenAIRE |
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