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Detection and identification of carcinogen-peptide adducts by nanoelectrospray tandem mass spectrometry

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Journal of the American Society for Mass Spectrometry

Abstract

Nanoelectrospray (nanoES) tandem mass spectrometry was used to examine covalently modified peptides in crude enzymatic digests of human serum albumin (HSA) that had been exposed to either benzo[a]pyrene diol epoxide (B[a]PDE, 1), chrysene diol epoxide (CDE, 2), 5-methylchrysene diol epoxide (5MeCDE, 3), or benzo[g]chrysene diol epoxide (B[g]CDE, 4). The low flow rates of nanoES (∼20 nL/min) allowed several MS/MS experiments to be optimized and performed on a single sample with very little sample consumption (∼30 min analysis time/µL sample). Initially, nanoES was compared with conventional LC/MS/MS analysis of carcinogen-peptide adducts. For example, nanoES analysis of an unseparated digest of B[a]PDE-treated serum albumin revealed the same peptides (RRHPY and RRHPY-FYAPE) that were previously shown, by LC/MS/MS, to be adducted with B[a]PDE. In addition, nanoES could detect unstable peptide adducts that might not otherwise have been directly observable. Finally, nanoES was shown to be an effective way to screen mixtures of modified and unmodified peptides for which no chromatographic information is available.

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References

  1. Calleman, C. J.; Ehrenberg, L.; Jansson, B.; Osterman-Golkar, S.; Segerbäck, D.; Sverisson, K.; Wachtneister, C. A. Monitoring and risk assessment by means of alkyl groups in hemoglobin in persons occupationally exposed to ethylene oxide. J. Environ. Pathol. Toxicol. 1978, 2, 427–444.

    CAS  Google Scholar 

  2. Ehrenberg, L.; Osterman-Golkar, S. Alkylation of macromolecules for detecting mutagenic agents. Teratog. Carcinog. Mutag. 1980, 1, 105–127.

    Article  CAS  Google Scholar 

  3. Osterman-Golkar, S.; Ehrenberg, L.; Segerbäck, D.; Hallstrom, I. Evaluation of genetic risks of alkylating agents. II. Haemoglobin as a dose monitor. Mutat. Res. 1976, 34, 1–10.

    CAS  Google Scholar 

  4. Osterman-Golkar, S.; Hultmark, D.; Segerbäck, D.; Calleman, C. J.; Gothe, R.; Ehrenberg, L.; Hallstrom, I. Alkylation of DNA and proteins in mice exposed to vinyl chloride. Biochem. Biophys. Res. Commun. 1977, 76, 259–266.

    Article  CAS  Google Scholar 

  5. Skipper, P. L.; Tannenbaum, S. R. Protein adducts in the molecular dosimetry of chemical carcinogens. Carcinogenesis 1990, 11, 507–518.

    Article  CAS  Google Scholar 

  6. Skipper, P. L.; Tannenbaum, S. R. Molecular dosimetry of aromatic amines in human populations. Environ. Health Persp. 1994, 102, Suppl. 17–21.

    Article  CAS  Google Scholar 

  7. Tannenbaum, S. R.; Skipper, P. L.; Wishnok, J. S.; Stillwell, W. G.; Day, B. W.; Taghizadeh, K. Characterization of various classes of protein adducts. Environ. Health Persp. 1993, 99, 51–55.

    Article  CAS  Google Scholar 

  8. Brunmark, P.; Harriman, S.; Skipper, P. L.; Wishnok, J. S.; Amin, S.; Tannenbaum, S. R. Identification of subdomain IB in human serum albumin as a major binding site for polycyclic aromatic hydrocarbon epoxides Chem. Res. Toxicol. 1997, 10, 880–886.

    Article  CAS  Google Scholar 

  9. Day, B. W.; Skipper, P. L.; Zaia, J.; Tannenbaum, S. R. Benzo[a]pyrene anti-diol epoxide covalently modifies human serum albumin carboxylate side chains and imidazole side chain of histidine(146). J. Am. Chem. Soc. 1991, 113, 8505–8509.

    Article  CAS  Google Scholar 

  10. Zaia, J.; Biemann, K. Characteristics of high energy collision-induced dissociation tandem mass spectra of polycyclic aromatic hydrocarbon diolepoxide adducted peptides. J. Am. Soc. Mass Spectrom. 1994, 5, 649–654.

    Article  CAS  Google Scholar 

  11. Wilm, M. S.; Mann, M. Electrospray and Taylor-Cone theory, Dole’s beam of macromolecules at last? Int. J. Mass Spectrom. Ion Proc. 1994, 136, 167–180.

    Article  CAS  Google Scholar 

  12. Wilm, M.; Mann, M. Analytical properties of the nanoelectrospray ion source. Anal. Chem. 1996, 68, 1–8.

    Article  CAS  Google Scholar 

  13. Wilm, M.; Shevchenko, A.; Houthaeve, T.; Brelt, S.; Schweigerer, L.; Fotsis, T.; Mann, M. Femtomole sequencing of proteins from polyacrylamide gels by nanoelectrospray mass spectrometry. Nature 1996, 379, 466–469.

    Article  CAS  Google Scholar 

  14. Shevchenko, A.; Wilm, M.; Vorm, O.; Mann, M. Mass spectrometric sequencing of proteins from silver-stained polyacrylamide gels. Anal. Chem. 1996, 68, 259–266.

    Article  Google Scholar 

  15. Davis, M. T.; Stahl, D. C.; Hefta, S. A.; Lee, T. D. A microscale electrospray interface for on-line, capillary liquid chromatography/tandem mass spectrometry of complex peptide mixtures. Anal. Chem. 1995, 67, 4549–4556.

    Article  CAS  Google Scholar 

  16. Korner, R.; Wilm, M.; Morand, K.; Schubert, M. Nano electrospray combined with a quadrupole ion trap for the analysis of peptides and protein digests. J. Am. Soc. Mass Spectrom. 1996, 7, 150–156.

    Article  Google Scholar 

  17. Valaskovic, G. A.; Kelleher, N. L.; Little, D. P.; Aasuerud, D. J.; McLafferty, F. W. Attomole sensitivity electrospray source for large-molecule mass spectrometry. Anal. Chem. 1995, 67, 3802–3805.

    Article  CAS  Google Scholar 

  18. Stryer, L. Biochemistry; W. H. Freeman:New Yok, 1988, pp 57–58.

    Google Scholar 

  19. Allen, G. , in Laboratory Techniques in Biochemistry and Molecular Biology; Burdon, R. H. , van Knippenberg, P. H. , Eds. ; Elsevier, New York, 1989, p 58.

  20. Baker, P. ; Clauser, K. MS-FIT. http://prospector. ucsf. edu/htmluscf/msfit-htm

  21. RamaKrishna, N. V. S.; Gao, F.; Padmavathi, N. S.; Cavalieri, E. L.; Rogan, E. G.; Cerny, R. L.; Gross, M. L. Model adducts of benzo[a]pyrene and nucleosides formed from its radical cation and diol epoxide Chem. Res. Toxicol. 1992, 5, 293–301.

    Article  CAS  Google Scholar 

  22. Szeliga, J.; Page, J. E.; Hilton, B. D.; Kiselyov, A. S.; Harvey, R. G.; Dunayevskiy, Y. M.; Vouros, P.; Dipple, A. Characterization of DNA adducts formed by anti-benzo[g]chrysene 11,12-dihydrodiol 13,14-epoxide Chem. Res. Toxicol. 1995, 8, 1014–1019.

    Article  CAS  Google Scholar 

  23. Annan, R. S.; Giese, R. W.; Vouros, P. Detection and structural characterization of amino polyaromatic hydrocarbon-de-oxynucleoside adducts using fast atom bombardment and tandem mass spectrometry Anal. Biochem. 1990, 191, 86–95

    Article  CAS  Google Scholar 

  24. Day, B. W.; Skipper, P. L.; Zaia, J.; Singh, K.; Tannenbaum, S. R. Enantiospecificity of covalent adduct formation by benzo[a]pyrene anti-diol epoxide with human serum albumin Chem. Res. Toxicol. 1994, 7, 829–835

    Article  CAS  Google Scholar 

  25. Day, B. W.; Skipper, P. L.; Rich, R. H.; Naylor, S.; Tannenbaum, S. R. Conversion of a hemoglobin alpha-chain aspartate(47) ester to N-(2,3-dihydroxypropyl)asparagine as a method for the identification of the principal binding site for benzo[a]pyrene anti-diol epoxide Chem. Res. Toxicol. 1991, 4, 359–363.

    Article  CAS  Google Scholar 

  26. Roepstorff, P.; Fohlman, J. Proposal for a common nomenclature for sequence ions in mass spectra of peptides Biomed. Mass Spectrom. 1984, 11, 601.

    Article  CAS  Google Scholar 

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Author notes

  1. James A. Hill

    Present address: Instrument Services, 28 Henderson Street, 02174, Arlington, MA

Authors and Affiliations

  1. Massachusetts Institute of Technology, Department of Chemistry, Cambridge, Massachusetts, USA

    Shawn P. Harriman, James A. Hill & Steven R. Tannenbaum

  2. Division of Toxicology, Massachusetts Institute of Technology, Room 56-747B, 02139, Cambridge, MA

    John S. Wishnok

Authors
  1. Shawn P. Harriman
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  2. James A. Hill
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  3. Steven R. Tannenbaum
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  4. John S. Wishnok
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Correspondence to John S. Wishnok.

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Harriman, S.P., Hill, J.A., Tannenbaum, S.R. et al. Detection and identification of carcinogen-peptide adducts by nanoelectrospray tandem mass spectrometry. J Am Soc Mass Spectrom 9, 202–207 (1998). https://doi.org/10.1016/S1044-0305(97)00252-3

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  • DOI: https://doi.org/10.1016/S1044-0305(97)00252-3

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