Česká společnost pro hmotnostní spektrometrii

(CEEPC/IPM/CMSC 2022 - ThO-05)
Advancing Cyclic Ion Mobility Mass Spectrometry Methods for Studying Biomolecules: Towards the Conformational Dynamics of Mega Dalton Protein Aggregates

Adam Pruška ^1 *, Julian Alexander Harrison ¹, Philipp Bittner ¹, Alexander Muck ², Dale A. Cooper-Shepherd ², Renato Zenobi ¹

1. ETH Zurich, Zurich, Switzerland
2. Waters Corporation, Wilmslow, Cheshire SK9 4AX, U.K.

Abstrakt

Native mass spectrometry is a powerful tool for the analysis of non-covalent complexes. When coupled with high-resolution ion mobility, this technique can be used to investigate the conformational changes induced in said complexes by different solution or gas-phase conditions. In this study, we describe how a new generation high-resolution ion mobility instrument equipped with a cyclic ion mobility cell can be utilized for the analysis of large biomolecular systems, including temperature-induced protein aggregates of masses greater than 1.5 MDa, as well as a 63 kDa oligonucleotide complex. The effects of and the interplay between the voltages applied to the different components of the cyclic ion mobility spectrometry system on ion transmission and arrival time distribution were demonstrated using biomolecules covering the m/z range 2 000 to 10 000. These data were used to establish a theoretical framework for achieving the best separation on the cyclic ion mobility system. Finally, the cyclic ion mobility mass spectrometer was coupled with a temperature-controlled electrospray ionization source to investigate high-mass protein aggregation. This analysis showed that it was possible to continuously monitor the change in abundance for several conformations of MDa aggregates with increasing temperature. This work significantly increases the range of biomolecules that can be analyzed by both cyclic ion mobility and temperature-controlled electrospray ionization mass spectrometry, providing new possibilities for high-resolution ion mobility analysis.

* Korespondující autor: pruskaadam@yahoo.co.uk

Literatura

1. Eldrid, C. et al. Anal. Chem. 91, 7554–7561 (2019).
2. Walker, T. E. et al. J Am Chem Soc. 144, 2667–2678.(2022).