Blackbody infrared radiative dissociation of nonspecific protein-carbohydrate complexes produced by nanoelectrospray ionization: The nature of the noncovalent interactions

  • Weijie Wang
  • Elena N. Kitova
  • Jiangxiao Sun
  • John S. Klassen
Articles

Abstract

Gas-phase thermal dissociation experiments, implemented with blackbody infrared radiative dissociation (BIRD) and Fourier transform ion cyclotron resonance mass spectrometry, have been performed on a series of protonated and deprotonated 1:1 and protonated 1:2 protein-carbohydrate complexes formed by nonspecific interactions during the nanoflow electrospray (nanoES) ionization process. Nonspecific interactions between the proteins bovine carbonic anhydrase II (CA), bovine ubiquitin (Ubq), and bovine pancreatic trypsin inhibitor and several carbohydrates, ranging in size from mono- to tetrasaccharides, have been investigated. Over the range of temperatures studied (60–190 °C), BIRD of the protonated and deprotonated complexes proceeds exclusively by the loss of the carbohydrate in its neutral form. The rates of dissociation of the 1:1 complexes containing a mono- or disaccharide decrease with reaction time, suggesting the presence of two or more kinetically distinct structures produced during nanoES or by gas-phase processes. In contrast, the 1:1 complexes of the tri- and tetrasaccharides exhibit simple first-order dissociation kinetics, a result that, on its own, is suggestive of a single preferred carbohydrate binding site or multiple equivalent sites in the gas phase. A comparative analysis of the dissociation kinetics measured for protonated 1:1 and 1:2 complexes of Ubq with αTal[αAbe]αMan further supports the presence of a single preferred binding site. However, a similar analysis performed on the complexes of CA and αTal[αAbe]αMan suggests that equivalent but dependent carbohydrate binding sites exist in the gas phase. Analysis of the Arrhenius activation parameters (Ea and A) determined for the dissociation of 1:1 complexes of CA with structurally related trisaccharides provides evidence that neutral intermolecular hydrogen bonds contribute, at least in part, to the stability of the gaseous complexes. Surprisingly, the Ea values for the complexes of the same charge state are not sensitive to the structure (primary or higher order) of the protein, suggesting that the carbohydrates are able to form energetically equivalent interactions with the various functional groups presented by the protein. For a given protein-carbohydrate complex, the dissociation Ea is sensitive to charge state, initially increasing and then decreasing with increasing charge. It is proposed that both ionic and neutral hydrogen bonds stabilize the nonspecific protein-carbohydrate complexes in the gas phase and that the relative contribution of the neutral and ionic interactions is strongly influenced by the charge state of the complex, with neutral interactions dominating at low charge states and ionic interactions dominating at high charge states.

References

  1. 1.
    Smith, D. L.; Zhang, Z. Probing Noncovalent Structural Features of Proteins by Mass Spectrometry. Mass Spectrom. Rev. 1994, 13, 411–429.CrossRefGoogle Scholar
  2. 2.
    Smith, R. D.; Bruce, J. E.; Wu, Q.; Lei, Q. P. New Mass Spectrometric Methods for the Study of Noncovalent Associations of Biopolymers. Chem. Soc. Rev. 1997, 26, 191–202.CrossRefGoogle Scholar
  3. 3.
    Loo, J. A. Electrospray Ionization Mass Spectrometry: A Technology for Studying Noncovalent Macromolecular Complexes. Int. J. Mass Spectrom. 2000, 200, 175–186.CrossRefGoogle Scholar
  4. 4.
    Hofstadler, S. A.; Griffey, R. H. Analysis of Noncovalent Complexes of DNA and RNA by Mass Spectrometry. Chem. Rev. 2001, 101, 377–390.CrossRefGoogle Scholar
  5. 5.
    Daniel, J. M.; Friess, S. D.; Rajagopalan, S.; Wendt, S.; Zenobi, R. Quantitative Determination of Noncovalent Binding Interactions Using Soft Ionization Mass Spectrometry. Int. J. Mass Spectrom. 2002, 216, 1–27.CrossRefGoogle Scholar
  6. 6.
    Heck, A. J. R.; van den Heuvel, R. H. H. Investigation of Intact Protein Complexes by Mass Spectrometry. Mass Spectrom. Rev. 2004, 23, 368–389.CrossRefGoogle Scholar
  7. 7.
    Li, Y. T.; Hsieh, Y. L.; Henion, J. D.; Senko, M. W.; McLafferty, F. W.; Ganem, B. Mass-Spectrometric Studies on Noncovalent Dimers of Leucine-Zipper Peptides. J. Am. Chem. Soc. 1993, 115, 8409–8413.CrossRefGoogle Scholar
  8. 8.
    Robinson, C. V.; Chung, E. W.; Kragelund, B. B.; Knudsen, J.; Aplin, R. T.; Poulsen, F. M.; Dobson, C. M. Probing the Nature of Noncovalent Interactions by Mass Spectrometry. A Study of Protein-CoA Ligand Binding and Assembly. J. Am. Chem. Soc. 1996, 118, 8646–8653.CrossRefGoogle Scholar
  9. 9.
    Wu, Q.; Gao, J.; Joseph-McCarthy, D.; Sigal, G. B.; Bruce, J. E.; Whitesides, G. M.; Smith, R. D. Carbonic Anhydrase-Inhibitor Binding: From Solution to the Gas Phase. J. Am. Chem. Soc. 1997, 119, 1157–1158.CrossRefGoogle Scholar
  10. 10.
    Kitova, E. N.; Bundle, D. R.; Klassen, J. S. Thermal Dissociation of Protein-Oligosaccharide Complexes in the Gas Phase: Mapping the Intrinsic Intermolecular Interactions. J. Am. Chem. Soc. 2002, 124, 5902–5913.CrossRefGoogle Scholar
  11. 11.
    Kitova, E. N.; Bundle, D. R.; Klassen, J. S. Evidence for the Preservation of Specific Intermolecular Interactions in Gaseous Protein-Oligosaccharide Complexes. J. Am. Chem. Soc. 2002, 124, 9340–9341.CrossRefGoogle Scholar
  12. 12.
    Felitsyn, N.; Peschke, M.; Kebarle, P. Origin and Number of Charges Observed on Multiply Protonated Native Proteins Produced by ESI. Int. J. Mass Spectrom. 2002, 219, 39–62.CrossRefGoogle Scholar
  13. 13.
    Carpenter, J. F.; Crowe, J. H. An Infrared Spectroscopic Study of the Interactions of Carbohydrates With Dried Proteins. Biochemistry 1989, 28, 3916–3922.CrossRefGoogle Scholar
  14. 14.
    Murray, B. S.; Liang, H.-J. Evidence for Conformational Stabilization of Beta-Lactoglobulin When Dried With Trehalose. Langmuir 2000, 16, 6061–6063.CrossRefGoogle Scholar
  15. 15.
    Wang, W.; Kitova, E. N.; Klassen, J. S. Bioactive Recognition Sites May Not Be Energetically Preferred in Protein-Carbohydrate Complexes in the Gas Phase. J. Am. Soc. Chem. 2003, 125, 13630–13631.CrossRefGoogle Scholar
  16. 16.
    Wang, W.; Kitova, E. N.; Klassen, J. S. Nonspecific Protein-Carbohydrate Complexes Produced by Nanoelectrospray Ionization. Factors Influencing Their Formation and Stability. Anal. Chem. 2005, 77, 3060–3071.CrossRefGoogle Scholar
  17. 17.
    Wang, W.; Kitova, E. N.; Klassen, J. S. Influence of Solution and Gas Phase Processes on Protein-Carbohydrate Binding Affinities Determined by Nanoelectrospray Fourier Transform Ion Cyclotron Resonance Mass Spectrometry. Anal. Chem. 2003, 75, 4945–4955.CrossRefGoogle Scholar
  18. 18.
    Felitsyn, N.; Kitova, E. N.; Klassen, J. S. Thermal Decomposition of a Gaseous Multiprotein Complex Studied by Blackbody Infrared Radiative Dissociation. Investigating the Origin of the Asymmetric Dissociation Behavior. Anal. Chem. 2001, 73, 4647–4661.CrossRefGoogle Scholar
  19. 19.
    Peschke, M.; Blades, A.; Kebarle, P. Charged States of Proteins. Reactions of Doubly Protonated Alkyldiamines With NH3: Solvation or Deprotonation. Extension of Two Proton Cases to Multiply Protonated Globular Proteins Observed in the Gas Phase. J. Am. Chem. Soc. 2002, 124, 11519–11530.CrossRefGoogle Scholar
  20. 20.
    Hunter, E. P. L.; Lias, S. G. Evaluated Gas Phase Basicities and Proton Affinities of Molecules: An Update. J. Phys. Chem. Ref. Data. 1998, 27, 413–656.CrossRefGoogle Scholar
  21. 21.
    Dunbar, R. C.; McMahon, T. B. Activation of Unimolecular Reactions by Ambient Blackbody Radiation. Science 1998, 279, 194–197.CrossRefGoogle Scholar
  22. 22.
    Price, W. D.; Williams, E. R. Activation of Peptide Ions by Blackbody Radiation: Factors That Lead to Dissociation Kinetics in the Rapid Energy Exchange Limit. J. Phys. Chem. A 1997, 101, 8844–8852.CrossRefGoogle Scholar
  23. 23.
    Price, W. D.; Williams, E. R. Binding Energies of the Proton-Bound Amino Acid Dimers Gly · Gly, Ala · Ala, Gly · Ala, and Lys · Lys Measured by Blackbody Infrared Radiative Dissociation. J. Phys. Chem. B 1997, 101, 664–673.CrossRefGoogle Scholar
  24. 24.
    Clemmer, D. E.; Hudgins, R. R.; Jarrold, M. F. Naked Protein Conformations—Cytochrome-c in the Gas-Phase. J. Am. Chem. Soc. 1995, 117, 10141–10142.CrossRefGoogle Scholar
  25. 25.
    Shelimov, K. B.; Jarrold, M. F. “Denaturation” and Refolding of Cytochrome c In Vacuo. J. Am. Chem. Soc. 1996, 118, 10313–10314.CrossRefGoogle Scholar
  26. 26.
    Shelimov, K. B.; Clemmer, D. E.; Hudgins, R. R.; Jarrold, M. F. Protein Structure In Vacuo: Gas-Phase Confirmations of BPTI and Cytochrome c. J. Am. Chem. Soc. 1997, 119, 2240–2248.CrossRefGoogle Scholar
  27. 27.
    Fye, J. L.; Woenckhaus, J.; Jarrold, M. F. Hydration of Folded and Unfolded Gas-Phase Proteins: Saturation of Cytochrome c and Apomyoglobin. J. Am. Chem. Soc. 1998, 120, 1327–1328.CrossRefGoogle Scholar
  28. 28.
    Li, J.; Taraszka, J. A.; Counterman, A. E.; Clemmer, D. E. Influence of Solvent Composition and Capillary Temperature on the Conformations of Electrosprayed Ions: Unfolding of Compact Ubiquitin Conformers From Pseudonative and Denatured Solutions. Int. J. Mass Spectrom. 1999, 185/186/187, 37–47.CrossRefGoogle Scholar
  29. 29.
    Mao, Y.; Ratner, M. A.; Jarrold, M. F. Molecular Dynamics Simulations of the Charge-Induced Unfolding and Refolding of Unsolvated Cytochrome c. J. Phys. Chem. B. 1999, 103, 10017–10021.CrossRefGoogle Scholar
  30. 30.
    Meot-Ner, M. The Ionic Hydrogen Bond and Ion Solvation. 2. Solvation of Onium Ions By One to Seven Water Molecules. Relations Between Monomolecular, Specific, and Bulk Hydrogen. J. Am. Chem. Soc. 1984, 106, 1265–1272.CrossRefGoogle Scholar
  31. 31.
    Meot-Ner, M. Ionic Hydrogen Bond and Ion Solvation. 6. Interaction Energies of the Acetate Ion With Organic Molecules. Comparison of CH3COO with Cl, CN, and SH. J. Am. Chem. Soc. 1988, 110, 3854–3858.CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2005

Authors and Affiliations

  • Weijie Wang
    • 1
  • Elena N. Kitova
    • 1
  • Jiangxiao Sun
    • 1
  • John S. Klassen
    • 1
  1. 1.Department of ChemistryUniversity of AlbertaEdmontonCanada

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