title: On the role of a direct interaction between protein ions and solvent additives during protein supercharging by electrospray ionization mass spectrometry.
abstract: The addition of certain reagents during the electrospray ionization mass spectrometry of proteins can shift the protein ion signal charge-state distributions (CSDs) to higher average charge states, a phenomenon known as 'supercharging'. The role of reagent gas-phase basicity (GB) during this process was investigated in both the negative and positive ion modes. Reagents with known or calculated GBs were added individually in equimolar amounts to protein solutions which were subsequently electrosprayed for mass spectrometry analysis. Shifts in the CSDs of the protein ion signals were monitored and related to the reagents' GBs. Trends for this data were evaluated for possible insights into a supercharging mechanism involving the direct interaction between supercharging reagent and protein ion. Reagent GB was confirmed to be directly related to the amount of supercharging observed in the negative ion mode. Supercharging in the positive ion mode, on the other hand, showed a maximal trend. Interestingly, a loss of signal and supercharging efficacy was observed for reagents with GBs intermediate within the investigated range, between ~800 and ~840 kJ mol(-1), at the 100 mM concentration used in the present study. The possibility of a direct interaction model for supercharging in the negative and positive ion modes dependent on the GBs of the protein ions and reagents is discussed. In the positive ion mode, supercharging appears to depend on the stability of a proton bridge formed between the reagent and a highly charged protein ion.
authors: Department of Chemistry, Western Michigan University, 1903 W Michigan Ave, Kalamazoo, MI 49008-5413, United States. kevin.douglass@mpiresearch.com.
authors: Kevin A
authors: KA
authors: Douglass
authors: D epartment of Chemistry, Western Michigan University, 1903 W Michigan Ave, Kalamazoo, MI 49008-5413, United States. andre.venter@wmich.edu.
authors: Andre R
authors: AR
authors: Venter