Abstract
Electrostatic interactions play an important role in the formation of noncovalent complexes. Our previous work has highlighted the role of certain amino acid residues, such as arginine, glutamate, aspartate, and phosphorylated/sulfated residues, in the formation of salt bridges resulting in noncovalent complexes between peptides. Tandem mass spectrometry (MS) studies of these complexes using collision-induced dissociation (CID) have provided information on their relative stability. However, product-ion spectra produced by CID have been unable to assign specifically the site of interaction for the complex. In this work, tandem MS experiments were conducted on noncovalent complexes using both electron capture dissociation (ECD) and electron-transfer dissociation (ETD). The resulting spectra were dominated by intramolecular fragments of the complex with the electrostatic interaction site intact. Based upon these data, we were able to assign the binding site for the peptides forming the noncovalent complex.
Article PDF
Avoid common mistakes on your manuscript.
References
O’Neil, K. T.; De Grado, W. F. A Predicted Structure of Calmodulin Suggests an Electrostatic Basis for its Function. Proc. Natl. Acad. Sci. U.S.A. 1985, 82, 4954–4958.
Woods, A. S.; Ciruela, F.; Fuxe, K.; Agnati, L. F.; Lluis, C.; Franco, R.; Ferre, S. Role of Electrostatic Interaction in Receptor-Receptor Heteromerization. J. Mol. Neurosci. 2005, 26, 125–132.
Kovacic, P.; Draskovich, C. D.; Pozos, R. S. Unifying Electrostatic Mechanism for Phosphates and Sulfates in Cell Signaling. J. Recept. Signal. Transduct. 2007, 27, 433–443.
Langner, M.; Kubica, K. The Electrostatics of Lipid Surfaces. Chem. Phys. Lipids 1999, 101, 3–35.
Johnson, L. N.; Lewis, R. J. Structural Basis for Control by Phosphorylation. Chem. Rev. 2001, 101, 2209–2242.
Schug, K. A.; Lindner, W. Noncovalent Binding between Guanidinium and Anionic Groups: Focus on Biological- and Synthetic-Based Arginine/Guanidinium and Interactions with Phosph[on]ate and Sulf[on]ate Residues. Chem. Rev. 2005, 105, 67–114.
Woods, A. S.; Huestis, M. A. A Study of Peptide—Peptide Interaction by Matrix-Assisted Laser Desorption/Ionization. J. Am. Soc. Mass Spectrom. 2001, 12, 88–96.
Jackson, S. N.; Wang, H.-Y. J.; Yergey, A.; Woods, A. S. Phosphate Stabilization of Intermolecular Interactions. J. Proteome Res. 2006, 5, 122–126.
Jackson, S. N.; Wang, H.-Y. J.; Woods, A. S. A Study of the Fragmentation Patterns of the Phosphate—Arginine Noncovalent Bond. J. Proteome Res. 2005, 4, 2360–2363.
Woods, A. S.; Wang, H.-Y. J.; Jackson, S. N. Sulfation, the Up-and-Coming Post-Translational Modification: Its Role and Mechanism in Protein—Protein Interaction. J. Proteome Res. 2007, 6, 1176–1182.
Zubarev, R. A.; Zubarev, A. R.; Savitski, M. M. Electron Capture/Transfer versus Collisionally Activated/Induced Dissociations: Solo or Duet? J. Am. Soc. Mass Spectrom. 2008, 19, 753–761.
Zubarev, R. A. Electron-Capture Dissociation Tandem Mass Spectrometry. Curr. Opin. Biotech. 2004, 15, 12–16.
Coon, J. J.; Syka, J. E. P.; Shabanowitz, J.; Hunt, D. F. Tandem Mass Spectrometry for Peptides and Protein Sequence Analysis. BioTechniques 2005, 38, 519–523.
Haselmann, K. F.; Jorgensen, T. J. D.; Budnik, B. A.; Jensen, F.; Zubarev, R. A. Electron Capture Dissociation of Weakly Bound Polypeptide Polycationic Complexes. Rapid Commun. Mass Spectrom. 2002, 16, 2260–2265.
Loo, J. A. Electrospray Ionization Mass Spectrometry: A Technology for Studying Noncovalent Macromolecular Complexes. Int. J. Mass Spectrom. 2000, 200, 175–186.
Author information
Authors and Affiliations
Corresponding author
Additional information
Published online September 6, 2008
Rights and permissions
About this article
Cite this article
Jackson, S.N., Dutta, S. & Woods, A.S. The use of ECD/ETD to identify the site of electrostatic interaction in noncovalent complexes. J Am Soc Mass Spectrom 20, 176–179 (2009). https://doi.org/10.1016/j.jasms.2008.08.021
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1016/j.jasms.2008.08.021