Supercharged protein and peptide ions formed by electrospray ionization

Analytical Chemistry
A T IavaroneE R Williams

Abstract

The multiple charging of large molecules in electrospray ionization provides key advantages for obtaining accurate molecular weights by mass spectrometry and for obtaining structural information by tandem mass spectrometry and MS(n) experiments. Addition of glycerol or m-nitrobenzyl alcohol into the electrospray solutions dramatically increases both the maximum observed charge state and the abundances of the high charge states of protein and peptide ions. Adding glycerol to acidified aqueous solutions of cytochrome c shifts the most abundant charge state from 17+ to 21+, shifts the maximum charge state from 20+ to 23+, and shifts the average charge state from 16.6+ to 20.9+. Much less m-nitrobenzyl alcohol (<1%) is required to produce similar results. With just 0.7% m-nitrobenzyl alcohol, even the 24+ charge state of cytochrome c is readily observed. Similar results are obtained with myoglobin and (Lys)4. For the latter molecule, the 5+ charge state is observed in the electrospray mass spectrum obtained from solutions containing 6.7% m-nitrobenzyl alcohol. This charge state corresponds to protonation of all basic sites in this peptide. Although the mechanism for enhanced charging is unclear, it does not appear to be a consequen...Continue Reading

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Citations

Apr 8, 2011·Journal of the American Society for Mass Spectrometry·Nadezda P SargaevaPeter B O'Connor
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Harry J SterlingEvan R Williams
Sep 29, 2011·Journal of the American Society for Mass Spectrometry·Lisa A VasicekJennifer S Brodbelt
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