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Multitechnique mass-spectrometric approach for the detection of bovine glutathione peroxidase selenoprotein: focus on the selenopeptide

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Abstract

Glutathione peroxidase (isolated from bovine erythrocytes) and its behaviour during alkylation and enzymatic digestion were studied by various hyphenated techniques: gel electrophoresis–laser ablation (LA) inductively coupled plasma (ICP) mass spectrometry (MS), size-exclusion liquid chromatography–ICP MS, capillary high-performance liquid chromatography (capHPLC)–ICP MS, matrix-assisted laser desorption/ionization (MALDI) time-of-flight (TOF) MS, electrospray MS, and nanoHPLC–electrospray ionization (ESI) MS/MS. ESI TOF MS and MALDI TOF MS allowed the determination of the molecular mass but could not confirm the presence of selenium in the protein. The purity of the protein with respect to selenium species could be evaluated by LA ICP MS and size-exclusion chromatography (SEC)–ICP MS under denaturating and nondenaturating conditions, respectively. SEC–ICP MS and capHPLC–ICP MS turned out to be valuable techniques to study the enzymolysis efficiency, miscleavage and artefact formation during derivatization and tryptic digestion. For the first time the parallel ICP MS and ESI MS/MS data are reported for the selenocysteine-containing peptide extracted from the gel; capHPLC–ICP MS allowed the sensitive detection of the selenopeptide regardless of the matrix and nanoHPLC–electrospray made possible its identification.

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References

  1. Rotruck JT, Pope AL, Ganther HE, Swanson AB, Hafeman DG, Hoekstra WG (1973) Science 179:588–590

    Article  CAS  Google Scholar 

  2. Flohe L, Günzler WA, Schock HH (1973) FEBS Letts 32:132–134

    Article  CAS  Google Scholar 

  3. Kryukov GV, Castellano S, Novoselov SV, Lobanov AV, Zehtab O, Guigó R, Gladyshev VN (2003) Science 300:1439–1443

    Article  CAS  Google Scholar 

  4. Epp O, Ladenstein R, Wendel A (1983) Eur J Biochem 133:51-69

    Article  CAS  Google Scholar 

  5. Chu FF, Doroshow JH, Esworhty RS (1993) J Biol Chem 268:2571–2576

    CAS  Google Scholar 

  6. Esworthy RS, Swiderek KM, Ho YS, Chu FF (1998) Biochim Biophys Acta 1381:213–226

    CAS  Google Scholar 

  7. Yoshimura S, Watanabe K, Suemizu H, Onozawa T, Mizoguchi J, Tsuda K, Hatta H, Moriuchi T (1991) J Biochem 109:918–923

    CAS  Google Scholar 

  8. Schuckelt R, Brigeliusflohe R, Maiorino M, Roveri A, Reumkens J, Strassburger W, Ursini F, Wolf B, Flohe L (1991) Free Radic Res Commun 14:343–361

    CAS  Google Scholar 

  9. Ursini F, Maiorino M, Gregolin C (1985) Biochim Biophys Acta 839:62–70

    CAS  Google Scholar 

  10. Gladyshev VN, Stadtman TC, Hatfield DL, Jeang K (1999) Proc Natl Acad Sci USA 96:835–839

    Article  CAS  Google Scholar 

  11. Jacobson GA, Narkowicz C, Tong YC, Peterson GM (2006) Clin Chim Acta 369:100–103

    Article  CAS  Google Scholar 

  12. Ma S, Caprioli RM, Hill KE, Burk RF (2003) J Am Soc Mass Spectrom 14:593–600

    Article  CAS  Google Scholar 

  13. Ghyselinck NB, Dufaure I, Lareyre JJ, Rigaudiere N, Mattei MG, Dufaure JP (1993) Mol Endocrinol 7:258–272

    Article  CAS  Google Scholar 

  14. Arthur JR (2000) Cel Mol Life Sci 57:1825–1835

    Article  CAS  Google Scholar 

  15. Lobinski R, Edmonds JS, Suzuki KT, Uden PC (2000) Pure Appl Chem 72:447–461

    CAS  Google Scholar 

  16. Koyama H, Kasanuma Y, Kim CY, Ejima A, Watanabe C, Satoh H (1996) Tohoku J Exp Med 178:17–25

    Article  CAS  Google Scholar 

  17. Koyama H, Omura K, Ejima A, Kasanuma Y, Watanabe C, Satoh H (1999) Anal Biochem 267:84–91

    Article  CAS  Google Scholar 

  18. Hinojosa-Reyes L, Marchante-Gayon JM, Alonso JIG, Sanz-Medel A (2003) J Anal At Spectrom 18:1210–1216

    Article  CAS  Google Scholar 

  19. Palacios O, Ruiz-Encinar J, Schaumloffel D, Lobinski R (2006) Anal Bioanal Chem 384:1276–1283

    Article  CAS  Google Scholar 

  20. Fan TWM, Pruszkowski E, Shuttleworth S (2002) J Anal At Spectrom 17:1621–1623

    Article  CAS  Google Scholar 

  21. Tastet L, Schaumloffel D, Bouyssiere B, Lobinski R (2006) Anal Bioanal Chem 385:948–953

    Article  CAS  Google Scholar 

  22. Ballihaut G, Tastet L, Pécheyran C, Bouyssiere B, Donard O, Grimaud R, Lobinski R (2005) J Anal At Spectrom 20:493–499

    Article  CAS  Google Scholar 

  23. Chery CC, Gunther D, Cornelis R, Vanhaecke F, Moens L (2003) Electrophoresis 24:3305–3313

    Article  CAS  Google Scholar 

  24. Chassaigne H, Chery CC, Bordin G, Rodriguez AR (2002) J Chromatogr A 976:409–422

    Article  CAS  Google Scholar 

  25. Qiu Y, Benet LZ, Burlingame AL (1998) J Biol Chem 273:17940–17953

    Article  CAS  Google Scholar 

  26. Mauri P, Benazzi L, Flohé L, Maiorino M, Pietta PG, Pilawa S, Roveri A, Ursini F (2003) Biol Chem 384:575–588

    Article  CAS  Google Scholar 

  27. Giusti P, Schaumlöffel D, Preud’homme H, Szpunar J, Łobiński R (2006) J Anal At Spectrom 21:26–32

    Article  CAS  Google Scholar 

  28. Dernovics M, Giusti P, Lobinski R (2007) J Anal At Spectrom 22:41–50

    Article  CAS  Google Scholar 

  29. Schaumloffël D, Ruiz-Encinar J, Lobinski R (2003) Anal Chem 75:6837–6842

    Article  Google Scholar 

  30. Gettins P, Dyal D, Crews B (1992) Arch Biochem Biophys 294:511–518

    Article  CAS  Google Scholar 

Download references

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Authors and Affiliations

  1. Laboratoire de Chimie Analytique Bio-inorganique et Environnement (UMR 5034), Hélioparc, 2, av. Pr. Angot, 64053, Pau, France

    Guillaume Ballihaut, Sandra Mounicou & Ryszard Lobinski

  2. Laboratoire d’Ecologie Moléculaire (Microbiologie), Université de Pau et des Pays de l’Adour, av. de l’Université, B.P. 1155, 64013, Pau, France

    Guillaume Ballihaut

  3. Department of Chemistry, Warsaw University of Technology, 00-664, Warszawa, Poland

    Ryszard Lobinski

Authors
  1. Guillaume Ballihaut
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  2. Sandra Mounicou
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  3. Ryszard Lobinski
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Corresponding author

Correspondence to Ryszard Lobinski.

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ESM 1

Sequences determined by nanoHPLC-electrospray MS/MS. The ionspresent in the collision induced dissociation mass spectra are shown either in green or in red (PPT 100 kb)

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Ballihaut, G., Mounicou, S. & Lobinski, R. Multitechnique mass-spectrometric approach for the detection of bovine glutathione peroxidase selenoprotein: focus on the selenopeptide. Anal Bioanal Chem 388, 585–591 (2007). https://doi.org/10.1007/s00216-007-1257-4

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  • DOI: https://doi.org/10.1007/s00216-007-1257-4

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