Česká společnost pro hmotnostní spektrometrii

(CEEPC/IPM/CMSC 2022 - FrO-10)
Two-dimensional mass spectrometry for top-down analysis and structural characterization of proteins

Maria van Agthoven ^1 *, Marek Polák ^1,2, Alan Kadek ¹, Michael Palasser ³, Marc-André Delsuc ^4,5, Kathrin Breuker ³, Petr Novák ^1,2

1. Institute of Microbiology of the Czech Academy of Sciences
2. Faculty of Science, Charles University
3. University of Innsbruck
4. IGBMC, Université de Strasbourg
5. CASC4DE

Abstrakt

Two-dimensional mass spectrometry (2DMS) is a method for tandem mass spectrometry that relies on ion radius modulation instead of ion isolation to correlate between precursor and fragment ion peaks. 2D mass spectra show all the fragmentation patterns of the analytes in a sample. Signal multiplexing yields high signal-to-noise ratios and therefore complete sequence coverage (e.g. for biomolecules) [1]. Modifications can easily be assigned and located visually with precursor ion scans and dissociation lines. 2DMS has also successfully been used for label-free relative quantification of modified histone peptides [2].
Acetylation is a covalent labelling method to probe tertiary and quaternary structures of proteins which has successfully been used in combination with top-down analysis to probe the structure of ubiquitin [3]. In this study, we analyse acetylated ubiquitin with 2DMS. We use the accuracy of the precursor-fragment correlation to identify and locate the acetylations in the sequence and we use fragment ion abundances for label-free relative quantification. We show that acetylation combined with 2DMS yields accurate information on ubiquitin tertiary structure.

* Korespondující autor: maria.vanagthoven@biomed.cas.cz

Literatura

1. van Agthoven M. et al.: Eur. Biophys. J. 48(3), 213-229 (2019).
2. Halper M. et al.: Anal. Chem. 92(20), 13945–13952 (2020),
3. Novak P. et al.: J. Mass Spectrom. 39, 322-328 (2004).

Poděkování:

The authors thank the International mobility of researchers of the Institute of Microbiology of the CAS, v.v.i. No 2, Number of project: CZ.02.2.69/0.0/0.0/18_053/0017705