Desalting large protein complexes during native electrospray mass spectrometry by addition of amino acids to the working solution

The Analyst
David J Clarke, Dominic J Campopiano

Abstract

Native mass spectrometry is a rapidly emerging field for characterising the structure of proteins and protein assemblies. The technique relies on electrospray ionisation (ESI) to efficiently ionise the protein analyte and transmit it into the gas phase with retention of protein structure, non-covalent protein-ligand and protein-protein interactions. In native ESI, both the ionisation efficiency and the resulting mass spectral signal is adversely effected by the presence of non-volatile inorganic salts, such as sodium chloride, which form extensive adducts with the protein ions. Consequently, there is great interest in finding experimental strategies that mitigate these phenomena. Here we report our findings that the addition of 10 mM l-serine to the ESI spray solution reduces the adverse effects of sodium adduction to proteins. In the analysis of bovine serum albumin (BSA; 66 kDa), 10 mM serine increased signal to noise ratio (S/N) ∼4 fold. This increase in sensitivity was accompanied by peak narrowing (∼10 fold), which allowed more precise assignment of molecular mass. Similar effects were observed when analysing protein complexes - serine palmitoyl transferase (SPT, a 92 kDa homodimer), enolase (a 93 kDa homodimer); and alcoh...Continue Reading

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Citations

Mar 21, 2017·Journal of the American Society for Mass Spectrometry·Elahe Honarvar, Andre R Venter
Jan 15, 2017·Proceedings of the National Academy of Sciences of the United States of America·Jean-Nicolas LongchampHans-Werner Fink
Jul 15, 2020·Journal of Mass Spectrometry : JMS·Marie Barth, Carla Schmidt
Feb 17, 2019·Journal of the American Society for Mass Spectrometry·Roshan JavanshadAndre R Venter
Oct 14, 2017·Journal of the American Society for Mass Spectrometry·Anna C SusaEvan R Williams
Jan 31, 2020·Journal of the American Society for Mass Spectrometry·Kazumi SaikusaSatoko Akashi
Feb 14, 2018·Analytical Chemistry·Zezhen ZhangR Graham Cooks
Aug 28, 2021·Chemical Reviews·Jack L BennettWilliam A Donald

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