Skip to main content
SpringerLink
Log in

Reagent Cluster Anions for Multiple Gas-Phase Covalent Modifications of Peptide and Protein Cations

  • Research Article
  • Published:
Journal of The American Society for Mass Spectrometry

Abstract

Multiple gas phase ion/ion covalent modifications of peptide and protein ions are demonstrated using cluster-type reagent anions of N-hydroxysulfosuccinimide acetate (sulfo-NHS acetate) and 2-formyl-benzenesulfonic acid (FBMSA). These reagents are used to selectively modify unprotonated primary amine functionalities of peptides and proteins. Multiple reactive reagent molecules can be present in a single cluster ion, which allows for multiple covalent modifications to be achieved in a single ion/ion encounter and at the ‘cost’ of only a single analyte charge. Multiple derivatizations are demonstrated when the number of available reactive sites on the analyte cation exceeds the number of reagent molecules in the anionic cluster (e.g., data shown here for reactions between the polypeptide [K10 + 3H]3+ and the reagent cluster [5R5Na – Na]–). This type of gas-phase ion chemistry is also applicable to whole protein ions. Here, ubiquitin was successfully modified using an FBMSA cluster anion which, upon collisional activation, produced fragment ions with various numbers of modifications. Data for the pentamer cluster are included as illustrative of the results obtained for the clusters comprised of two to six reagent molecules.

ᅟ

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Figure 1
Figure 2
Figure 3
Figure 4
Figure 5
Figure 6

Similar content being viewed by others

References

  1. Biemann, K.: Mass spectrometry of peptides and proteins. Annu. Rev. Biochem. 61, 977–1010 (1992)

    Article  CAS  Google Scholar 

  2. Halket, J.M., Zaikin, V.G.: Derivatization in mass spectrometry – 1. Silylation. Eur. J. Mass Spectrom. 9, 1–21 (2003)

    Article  CAS  Google Scholar 

  3. Hermanson, G.T.: Bioconjugate Techniques, 2nd edn. Academic Press, Amsterdam (1998)

    Google Scholar 

  4. Yang, W.C., Mirzaei, H., Liu, X.P., Regnier, R.E.: Enhancement of amino acid detection and quantification by electrospray ionization mass spectrometry. Anal. Chem. 78, 4702–4708 (2006)

    Article  CAS  Google Scholar 

  5. Gygi, S.P., Rist, B., Gerber, S.A., Turecek, F., Gelb, M.H., Aebersold, R.: Quantitative analysis of complex protein mixtures using isotope-coded affinity tags. Nat. Biotechnol. 17, 994–999 (1999)

    Article  CAS  Google Scholar 

  6. Ross, P.L., Huang, Y.N., Marchese, J.N., Williamson, B., Parker, K., Hattan, S., Khainovski, N., Pillai, S., Dey, S., Daniels, S., Purkayastha, S., Juhasz, P., Martin, S., Bartlet-Jones, M., He, F., Jacobson, A., Pappin, D.J.: Multiplexed protein quantitation in Saccharomyces cerevisiae using amine-reactive isobaric tagging reagents. Mol. Cell Proteomics 3, 1154–1169 (2004)

    Article  CAS  Google Scholar 

  7. Beardsley, R.L., Sharon, L.A., Reilly, J.P.: Peptide de novo sequencing facilitated by a dual-labeling strategy. Anal. Chem. 77, 6300–6309 (2005)

    Article  CAS  Google Scholar 

  8. Madsen, J.A., Brodbelt, J.S.: Simplifying fragmentation patterns of multiply charged peptides by N-terminal derivatization and electron transfer collision activated dissociation. Anal. Chem. 81, 3645–3653 (2009)

    Article  CAS  Google Scholar 

  9. Novak, P., Kruppa, G.H., Young, M.M., Schoeniger, J.: A top-down method for the determination of residue-specific solvent accessibility in proteins. J. Mass Spectrom. 39, 322–328 (2004)

    Article  CAS  Google Scholar 

  10. Tubb, M.R., Silva, R.A.G.D., Fang, J., Tso, P., Davidson, W.S.: Lipids and lipoproteins: metabolism, regulation, and signaling. J. Biol. Chem. 283, 17314–17323 (2008)

    Article  CAS  Google Scholar 

  11. Singh, P., Panchaud, A., Goodlett, D.R.: Chemical cross-linking and mass spectrometry as a low-resolution protein structure determination technique. Anal. Chem. 82, 2636–2642 (2010)

    Article  CAS  Google Scholar 

  12. Chen, Z., Jawhari, A., Fischer, L., Buchen, C., Tahir, S., Kamenski, T., Rasmussen, M., Lariviere, L., Bukowsk-Willis, J.C., Nilges, M., Cramer, P., Rappsilber, J.: Architecture of the RNA polymerase II–TFIIF complex revealed by cross-linking and mass spectrometry. EMBO J. 29, 717–726 (2010)

    Article  CAS  Google Scholar 

  13. Vasicek, L., Brodbelt, J.S.: Enhancement of ultraviolet photodissociation efficiencies through attachment of aromatic chromophores. Anal. Chem. 82, 9441–9446 (2010)

    Article  CAS  Google Scholar 

  14. Venter, A., Sojka, P.E., Cooks, R.G.: Droplet dynamics and ionization mechanisms in desorption electrospray ionization mass spectrometry. Anal. Chem. 78, 8549–8555 (2006)

    Article  CAS  Google Scholar 

  15. Girod, M., Moyano, E., Campbell, D.I., Cooks, R.G.: Accelerated bimolecular reactions in microdroplets studied by desorption electrospray ionization mass spectrometry. Chem. Sci. 2, 501–510 (2011)

    Article  CAS  Google Scholar 

  16. Miao, Z., Chen, H.: Direct analysis of liquid samples by desorption electrospray ionization-mass spectrometry (DESI-MS). J. Am. Soc. Mass Spectrom. 20, 10–19 (2009)

    Article  CAS  Google Scholar 

  17. Perry, R.H., Splendore, M., Chien, A., Davis, N.K., Zare, R.N.: Detecting reaction intermediates in liquids on the millisecond time scale using desorption electrospray ionization. Angew. Chem. Int. Ed. 50, 250–254 (2011)

    Article  CAS  Google Scholar 

  18. Badu-Tawiah, A.K., Li, A., Jjunju, F.P.M., Cooks, R.G.: Peptide cross-linking at ambient surfaces by reactions of nanosprayed molecular cations. Angew. Chem. Int. Ed. 51, 9417–9421 (2012)

    Article  CAS  Google Scholar 

  19. Prentice, B.M., McLuckey, S.A.: Gas-phase ion/ion reactions of peptides and proteins: acid/base, redox, and covalent chemistries. Chem. Commun. 49, 935–942 (2013)

    Article  Google Scholar 

  20. Mentinova, M., McLuckey, S.A.: Intra- and intermolecular cross-linking of peptide ions in the gas-phase: reagents and conditions. J. Am. Soc. Mass Spectrom. 22, 912–921 (2011)

    Article  CAS  Google Scholar 

  21. Webb, I.K., Mentinova, M., McGee, W.M., McLuckey, S.A.: Gas-phase intramolecular protein crosslinking via ion/ion reactions: ubiquitin and a homobifunctional sulfo-NHS ester. J. Am. Soc. Mass Spectrom. doi:10.1007/s13361-013-0590-4 (2013)

  22. Mentinova, M., McLuckey, S.A.: Covalent modification of gaseous peptide ions with N-hydroxysuccinimide ester reagent ions. J. Am. Chem. Soc. 132, 18248–18257 (2010)

    Article  CAS  Google Scholar 

  23. Mentinova, M., Barefoot, N.Z., McLuckey, S.A.: Solution versus gas-phase modification of peptide cations with NHS-ester reagents. J. Am. Soc. Mass Spectrom. 23, 282–289 (2011)

    Article  Google Scholar 

  24. McGee, W.M., Mentinova, M., McLuckey, S.A.: Gas-phase conjugation to arginine residues in polypeptide ions via N-hydroxysuccinimide ester-based reagent ions. J. Am. Chem. Soc. 134, 11412–11414 (2012)

    Article  CAS  Google Scholar 

  25. Han, H., McLuckey, S.A.: Selective covalent bond formation in polypeptide ions via gas-phase ion/ion reaction chemistry. J. Am. Chem. Soc. 131, 12884–12885 (2009)

    Article  CAS  Google Scholar 

  26. Hassell, K.M., Stutzman, J.R., McLuckey, S.A.: Gas-phase bioconjugation of peptides via ion/ion charge inversion: Schiff base formation on the conversion of cations to anions. Anal. Chem. 82, 1594–1597 (2010)

    Article  CAS  Google Scholar 

  27. Stutzman, J.R., Hassel, K.M., McLuckey, S.A.: Dissociation behavior of tryptic and intramolecular disulfide-linked peptide ions modified in the gas phase via ion/ion reactions. Int. J. Mass Spectrom. 312, 195–200 (2012)

    Article  CAS  Google Scholar 

  28. Stutzman, J.R., McLuckey, S.A.: Ion/ion reactions of MALDI-derived peptide ions: increased sequence coverage via covalent and electrostatic modification upon charge inversion. Anal. Chem. 84, 10679–10685 (2012)

    Article  CAS  Google Scholar 

  29. Prentice, B.M., Gilbert, J.D., Stutzman, J.R., Forrest, W.P., McLuckey, S.A.: Gas-phase reactivity of carboxylic acid functional groups with carbodiimides. J. Am. Soc. Mass Spectrom. 24, 30–37 (2013)

    Article  CAS  Google Scholar 

  30. Gillen, G., Roberson, S.: Preliminary evaluation of an SF5+ polyatomic primary ion beam for analysis of organic thin films by secondary ion mass spectrometry. Rapid Commun. Mass Spectrom. 12, 1303–1312 (1998)

    Article  CAS  Google Scholar 

  31. Mahoney, C.M.: Cluster secondary ion mass spectrometry of polymers and related materials. Mass Spectrom. Rev. 29, 247–293 (2010)

    CAS  Google Scholar 

  32. McLuckey, S.A., Van Berkel, G.J., Glish, G.L.: Reactions of dimethylamine with multiply charged ions of cytochrome c. J. Am. Chem. Soc. 112, 5668–5670 (1990)

    Article  CAS  Google Scholar 

  33. McLuckey, S.A., Glish, G.L., Van Berkel, G.J.: Charge determination of product ions formed from collision-induced dissociation of multiply protonated molecules via ion/molecule reactions. Anal. Chem. 63, 1971–1978 (1991)

    Article  CAS  Google Scholar 

  34. Brutschy, B.: Ion–molecule reactions within molecular clusters. Chem. Rev. 92, 1567–1587 (1992)

    Article  CAS  Google Scholar 

  35. Spangler, G.E.: Fundamental considerations for the application of miniature ion mobility spectrometry to field analytical applications. Field Anal. Chem. Technol. 4, 255–267 (2000)

    Article  CAS  Google Scholar 

  36. Krylova, N., Krylov, E., Eiceman, G.A., Stone, J.A.: Effect of moisture on the field dependence of mobility for gas-phase ions of organophosphorus compounds at atmospheric pressure with field asymmetric ion mobility spectrometry. J. Phys. Chem. A 107, 3648–3654 (2003)

    Article  CAS  Google Scholar 

  37. Bruins, A.P., Covey, T.R., Henion, J.D.: Ion spray interface for combined liquid chromatography/atmospheric pressure ionization mass spectrometry. Anal. Chem. 59, 2642–2646 (1987)

    Article  CAS  Google Scholar 

  38. Chowdhury, S.K., Katta, V., Chait, B.T.: An electrospray-ionization mass spectrometer with new features. Rapid Commun. Mass Spectrom. 4, 81–87 (1990)

    Article  CAS  Google Scholar 

  39. He, M., McLuckey, S.A.: Two ion/ion charge inversion steps to form a doubly protonated peptide from a singly protonated peptide in the gas phase. J. Am. Chem. Soc. 125, 7756–7757 (2003)

    Article  CAS  Google Scholar 

  40. He, M., McLuckey, S.A.: Increasing the negative charge of a macroanion in the gas phase via sequential charge inversion reactions. Anal. Chem. 76, 4189–4192 (2004)

    Google Scholar 

  41. He, M., Emory, J.F., McLuckey, S.A.: Reagent anions for charge inversion of polypeptide/protein cations in the gas phase. Anal. Chem. 77, 3173–3182 (2005)

    Article  CAS  Google Scholar 

  42. Gunawardena, H.P., Emory, J.F., McLuckey, S.A.: Phosphopeptide anion characterization via sequential charge inversion and electron-transfer dissociation. Anal. Chem. 78, 3788–3793 (2006)

    Article  CAS  Google Scholar 

  43. Emory, J.F., McLuckey, S.A.: Charge inversion of polypeptide anions using protein and dendrimer cations as charge inversion reagents. Int. J. Mass Spectrom. 276, 102–109 (2008)

    Article  CAS  Google Scholar 

  44. Emory, J.F., McLuckey, S.A.: The role of amino acid composition in the charge inversion of deprotonated peptides via gas-phase ion/ion reactions. J. Am. Soc. Mass Spectrom. 20, 180–187 (2009)

    Article  CAS  Google Scholar 

  45. Hassell, K., LeBlanc, Y., McLuckey, S.A.: Conversion of multiple analyte cation types to a single analyte anion type via ion/ion charge inversion. Analyst 134, 2262–2266 (2009)

    Article  CAS  Google Scholar 

  46. Hassell, K., LeBlanc, Y., McLuckey, S.A.: Chemical noise reduction via mass spectrometry and ion/ion charge inversion: amino acids. Anal. Chem. 83, 3252–3255 (2011)

    Article  CAS  Google Scholar 

  47. Bower, J.J., Hodges, B.D.M., Saad, O.M., Leary, J.A., McLuckey, S.A.: Proton hydrates as soft ion/ion proton transfer reagents for multiply deprotonated biomolecules. Int. J. Mass Spectrom. 276, 153–159 (2008)

    Article  Google Scholar 

  48. Xia, Y., Wu, J., Londry, F.A., Hager, J.W., McLuckey, S.A.: Mutual storage mode ion/ion reactions in a hybrid linear ion trap. J. Am. Soc. Mass Spectrom. 16, 71–81 (2005)

    Article  CAS  Google Scholar 

  49. Liang, X., Xia, Y., McLuckey, S.A.: Alternately pulsed nano-electrospray ionization/atmospheric pressure chemical ionization for ion/ion reactions in an electrodynamic ion trap. Anal. Chem. 78, 3208–3212 (2006)

    Article  CAS  Google Scholar 

  50. Liang, X., Han, H., Xia, Y., McLuckey, S.A.: A pulsed triple ionization source for sequential ion/ion reactions in an electrodynamic ion trap. J. Am. Soc. Mass Spectrom. 18, 369–376 (2007)

    Article  CAS  Google Scholar 

  51. Londry, F.A., Hager, J.W.: Mass selective axial ion ejection from a linear quadrupole ion trap. J. Am. Soc. Mass Spectrom. 14, 1130–1147 (2003)

    Article  CAS  Google Scholar 

  52. Kish, M.M., Ohanessian, H., Wesdemiotis, C.: The Na+ affinities of α-amino acids: side-chain substituent effects. Int. J. Mass Spectrom. 227, 509–524 (2003)

    Article  CAS  Google Scholar 

  53. House Jr., J.E., Kemper, K.A.: Proton affinities of sulfate and bisulfate ions. J. Therm. Anal. 32, 1855–1858 (1987)

    Article  CAS  Google Scholar 

  54. Remko, M., Van Duijnen, P.T., von der Lieth, C.: Structure and stability of Li(I) and Na(I)- carboxylate, sulfate and phosphate complexes. J. Mol. Struct. (THEOCHEM) 814, 119–125 (2007)

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors acknowledge support for this work by AB SCIEX and by the National Institutes of Health under grant GM 45372. B.M.P. acknowledges receipt of a Purdue University Bilsland Dissertation Fellowship.

Author information

Authors and Affiliations

  1. Department of Chemistry, Purdue University, West Lafayette, IN, 47907-2084, USA

    Boone M. Prentice, John R. Stutzman & Scott A. McLuckey

Authors
  1. Boone M. Prentice
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John R. Stutzman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Scott A. McLuckey
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Scott A. McLuckey.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Prentice, B.M., Stutzman, J.R. & McLuckey, S.A. Reagent Cluster Anions for Multiple Gas-Phase Covalent Modifications of Peptide and Protein Cations. J. Am. Soc. Mass Spectrom. 24, 1045–1052 (2013). https://doi.org/10.1007/s13361-013-0637-6

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13361-013-0637-6

Key words

Search

Navigation