Probing the hydrophobic effect of noncovalent complexes by mass spectrometry

  • Claudia Bich
  • Samuel Baer
  • Matthias C. Jecklin
  • Renato Zenobi
Short Communication

Abstract

The study of noncovalent interactions by mass spectrometry has become an active field of research in recent years. The role of the different noncovalent intermolecular forces is not yet fully understood since they tend to be modulated upon transfer into the gas phase. The hydrophobic effect, which plays a major role in protein folding, adhesion of lipid bilayers, etc., is absent in the gas phase. Here, noncovalent complexes with different types of interaction forces were investigated by mass spectrometry and compared with the complex present in solution. Creatine kinase (CK), glutathione S-transferase (GST), ribonuclease S (RNase S), and leucine zipper (LZ), which have dissociation constants in the nM range, were studied by native nanoelectrospray mass spectrometry (nanoESI-MS) and matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS) combined with chemical cross-linking (XL). Complexes interacting with hydrogen bonds survived the transfer into gas phase intact and were observed by nanoESI-MS. Complexes that are bound largely by the hydrophobic effect in solution were not detected or only at very low intensity. Complexes with mixed polar and hydrophobic interactions were detected by nanoESI-MS, most likely due to the contribution from polar interactions. All noncovalent complexes could easily be studied by XL MALDI-MS, which demonstrates that the noncovalently bound complexes are conserved, and a real “snap-shot” of the situation in solution can be obtained.

Keywords

MALDI Leucine Zipper Electron Capture Dissociation Noncovalent Interaction Polar Interaction 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Loo, J. A. Studying Noncovalent Protein Complexes by Electrospray Ionization Mass Spectrometry. Mass Spectrom. Rev. 1997, 16, 1–23.CrossRefGoogle Scholar
  2. 2.
    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
  3. 3.
    Berg, J.; Tymoczko, J.; Stryer, L. Biochemistry 5th ed. 2002, Series Michelle Julet; Freeman: New York, pp 43–51.Google Scholar
  4. 4.
    Southall, N. T.; Dill, K. A.; Haymet, A. D. J. A View of the Hydrophobic Effect. J. Phys. Chem. B 2002, 106, 521–533.CrossRefGoogle Scholar
  5. 5.
    Pace, C. N.; Shirley, B. A.; McNutt, M.; Gajiwala, K. Forces Contributing to the Conformational Stability of Proteins. FASEB J. 1996, 10, 75–83.Google Scholar
  6. 6.
    Ball, P. Water as an Active Constituent in Cell Biology. Chem. Rev. 2008, 108, 74–108.CrossRefGoogle Scholar
  7. 7.
    Widom, B.; Bhimalapuram, P.; Koga, K. The Hydrophobic Effect. Phys. Chem. Chem. Phys. 2003, 5, 3085–3093.CrossRefGoogle Scholar
  8. 8.
    Kauzmann, W. Some Factors in the Interpretation of Protein Denaturation. Adv. Protein Chem. 1959, 14, 1–63.CrossRefGoogle Scholar
  9. 9.
    Breuker, K.; McLafferty, F. W. The Thermal Unfolding of Native Cytochrome c in the Transition from Solution to Gas Phase Probed by Native Electron Capture Dissociation. Angew. Chem. Int. Ed. 2005, 44, 4911–4914.CrossRefGoogle Scholar
  10. 10.
    Patriksson, A.; Marklund, E.; van der Spoel, D. Protein Structures Under Electrospray Conditions. Biochemistry 2007, 46, 933–945.CrossRefGoogle Scholar
  11. 11.
    Laskin, J.; Futrell, J. H. Entropy is the Major Driving Force for Fragmentation of Proteins and Protein—Ligand Complexes in the Gas Phase. J. Phys. Chem. A 2003, 107, 5836–5839.CrossRefGoogle Scholar
  12. 12.
    Franski, R.; Gierczyk, B.; Schroeder, G.; Franska, M.; Wyrwas, B. Do Hydrophobic Interactions Exist in the Gas Phase? Rapid Commun. Mass Spectrom. 2008, 22, 1339–1343.CrossRefGoogle Scholar
  13. 13.
    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
  14. 14.
    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
  15. 15.
    Bovet, C.; Wortmann, A.; Eiler, S.; Granger, F.; Ruff, M.; Gerrits, B.; Moras, D.; Zenobi, R. Estrogen Receptor—Ligand Complexes Measured by Chip-Based Nanoelectrospray Mass Spectrometry: An Approach for the Screening of Endocrine Disruptors. Protein Sci. 2007, 16, 938–946.CrossRefGoogle Scholar
  16. 16.
    Yin, S.; Xie, Y.; Loo, J. A. Mass Spectrometry of Protein—Ligand Complexes: Enhanced Gas-Phase Stability of Ribonuclease—Nucleotide Complexes. J. Am. Soc. Mass Spectrom. 2008, 19, 1199–1208.CrossRefGoogle Scholar
  17. 17.
    Li, Y.; Heitz, F.; Le Grimellec, C.; Cole, R. B. Hydrophobic Component in Noncovalent Binding of Fusion Peptides to Lipids as Observed by Electrospray Mass Spectrometry. Rapid Commun. Mass Spectrom. 2004, 18, 135–137.CrossRefGoogle Scholar
  18. 18.
    Goodlett, D. R.; Ogorzalek Loo, R.; Loo, J. A.; Wahl, J. H.; Udseth, H. R.; Smith, R. D. A Study of the Thermal Denaturation of Ribonuclease S by Electrospray Ionization Mass Spectrometry. J. Am. Soc. Mass Spectrom. 1994, 5, 614–622.CrossRefGoogle Scholar
  19. 19.
    Liang, Y.; Du, F.; Sanglier, S.; Zhou, B.; Xia, Y.; Van Dorsselaer, A.; Maechling, C.; Kilhoffer, M.; Haiech, J. J. Unfolding of Rabbit Muscle Creatine Kinase Induced by Acid. J. Biol. Chem. 2003, 278, 30098–30105.CrossRefGoogle Scholar
  20. 20.
    Tahallah, N.; Pinkse, M.; Maier, C. S.; Heck, A. J. The Effect of the Source Pressure on the Abundance of Ions of Noncovalent Protein Assemblies in an Electrospray Ionization Orthogonal Time-of-Flight Instrument. Rapid Commun. Mass Spectrom. 2001, 15, 596–601.CrossRefGoogle Scholar
  21. 21.
    Strupat, K.; Sagi, D.; Bonisch, H.; Schafer, G.; Peter-Katalinic, J. Oligomerization and Substrate Binding Studies of the Adenylate Kinase from Sulfolobus Acidocaldarius by Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry. Analyst 2000, 125, 563–567.CrossRefGoogle Scholar
  22. 22.
    Friess, S. D.; Daniel, J. M.; Zenobi, R. Probing the Surface Accessibility of Proteins with Noncovalent Receptors and MALDI Mass Spectrometry. Phys. Chem., Chem. Phys. 2004, 6, 2664–2675.CrossRefGoogle Scholar
  23. 23.
    Farmer, T. B.; Caprioli, R. M. Assessing the Multimeric States of Proteins-Studies Using Laser Desorption Mass-Spectrometry. Biol. Mass Spectrom. 1991, 20, 796–800.CrossRefGoogle Scholar
  24. 24.
    Bich, C.; Madler, S.; Chiesa, K.; DeGiacomo, F.; Bogliotti, N.; Zenobi, R. Reactivity and Applications of New Amino Reactive Cross-Linkers for Mass Spectrometric Detection of Protein—Protein Complexes. Anal. Chem. 2009, unpublished (submitted).Google Scholar
  25. 25.
    Cox, J. M.; Chan, C. A.; Chan, C.; Jourden, M. J.; Jorjorian, A. D.; Brym, M. J.; Snider, M. J.; Borders, C. L.; Edmiston, P. L. Generation of an Active Monomer of Rabbit Muscle Creatine Kinase by Site-Directed Mutagenesis: The Effect of Quaternary Structure on Catalysis and Stability. Biochemistry 2003, 42, 1863–1871.CrossRefGoogle Scholar
  26. 26.
    Rao, J. K.; Bujacz, G.; Wlodawer, A. Crystal Structure of Rabbit Muscle Creatine Kinase. FEBS Lett. 1998, 439, 133–137.CrossRefGoogle Scholar
  27. 27.
    Mannervik, B.; Jensson, H. Binary Combinations of Four Protein Subunits with Different Catalytic Specificities Explain the Relationship Between 6 Basic Glutathione S-Transferases in Rat-Liver Cytosol. J. Biol. Chem. 1982, 257, 9909–9912.Google Scholar
  28. 28.
    Hornby, J. A. T.; Codreanu, S. G.; Armstrong, R. N.; Dirr, H. W. Molecular Recognition at the Dimer Interface of a Class Mu Glutathione Transferase: Role of a Hydrophobic Interaction Motif in Dimer Stability and Protein Function. Biochemistry 2002, 41, 14238–14247.CrossRefGoogle Scholar
  29. 29.
    Wallace, L. A.; Dirr, H. Folding and Assembly of Dimeric Human Glutathione Transferase A1-1. Biochemistry 1999, 38, 16686–16694.CrossRefGoogle Scholar
  30. 30.
    Loo, R. R. O.; Goodlett, D. R.; Smith, R. D.; Loo, J. A. Observation of a Noncovalent Ribonuclease S-Protein S-Peptide Complex by Electrospray Ionization Mass-Spectrometry. J. Am. Chem. Soc. 1993, 115, 4391–4392.CrossRefGoogle Scholar
  31. 31.
    Madler, S.; Bich, C.; Touboul, D.; Zenobi, R. Chemical Crosslinking with NHS Esters: A Systematic Study on Amino Acid Reactivities. J. Mass Spectrom. 2009, 44, 694–706.CrossRefGoogle Scholar
  32. 32.
    Moitra, J.; Szilak, L.; Krylov, D.; Vinson, C. Leucine is the Most Stabilizing Aliphatic Amino Acid in the D Position of a Dimeric Leucine Zipper Coiled Coil. Biochemistry 1997, 36, 12567–12573.CrossRefGoogle Scholar
  33. 33.
    Wendt, H.; Baici, A.; Bosshard, H. R. Mechanism of Assembly of a Leucine-Zipper Domain. J. Am. Chem. Soc. 1994, 116, 6973–6974.CrossRefGoogle Scholar
  34. 34.
    Wendt, H.; Durr, E.; Thomas, R. M.; Przybylski, M.; Bosshard, H. R. Characterization of Leucine-Zipper Complexes by Electrospray-Ionization Mass-Spectrometry. Protein Sci. 1995, 4, 1563–1570.CrossRefGoogle Scholar

Copyright information

© American Society for Mass Spectrometry 2010

Authors and Affiliations

  • Claudia Bich
    • 1
  • Samuel Baer
    • 1
  • Matthias C. Jecklin
    • 1
  • Renato Zenobi
    • 1
  1. 1.Department of Chemistry and Applied BiosciencesETH ZurichZurichSwitzerland

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