Some critical aspects in the determination of binding constants by electrospray ionisation mass spectrometry at the example of arsenic bindings to sulphur-containing biomolecules

Journal of Mass Spectrometry : JMS
Anne-Christine Schmidt, Sandra Steier

Abstract

The influences of reactant concentrations, solvent type, acid strength, pH conditions and ionic strength on the determination of apparent gas-phase equilibrium constants K using electrospray ionisation mass spectrometry (ESI-MS) were elucidated. As example serves the interaction of the tripeptide glutathione (GSH) with phenylarsine oxide (PAO). It was shown that rising initial concentrations of both reactants were not adequately compensated by increasing signal intensities of the reaction products in the mass spectra. The equilibrium constant for the formation of the phenylarsenic-substituted peptide species decreased from 1.42 x 10(5) +/- 1.81 x 10(4) l micromol(-1) to 1.54 x 10(4) +/- 1.5 x 10(3) l micromol(-1) with rising initial GSH concentrations from 1 to 10 microM at fixed PAO molarity of 50 microM. K values resulting from a series with a fixed GSH molarity of 5 microM and a PAO molarity varied from 10 to 100 microM remained in a narrower range between 4.59 x 10(4) +/- 2.15 x 10(4) l micromol(-1) and 1.07 x 10(4) +/- 4.0 x 10(3) l micromol(-1). In contrast, consumption numbers calculated from the ion intensity ratios of reaction products to the unreacted peptide were not influenced by the initial reactant concentrations....Continue Reading

References

May 15, 2003·Journal of the American Society for Mass Spectrometry·Jürg M DanielRenato Zenobi
Oct 8, 2003·Journal of the American Society for Mass Spectrometry·Yannis Dotsikas, Yannis L Loukas
Dec 4, 2003·Journal of the American Society for Mass Spectrometry·Silke WendtRenato Zenobi
Nov 16, 2004·Rapid Communications in Mass Spectrometry : RCM·Kris De VriendtBart Devreese
Dec 13, 2006·Rapid Communications in Mass Spectrometry : RCM·Anne-Christine SchmidtMatthias Otto
Oct 9, 2007·Rapid Communications in Mass Spectrometry : RCM·Christopher T Veros, Neil J Oldham
Jul 12, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Ruben t'KindtJan Van Bocxlaer

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