Skip to main content

Advertisement

SpringerLink
Log in

Differential proteome analysis of tonsils from children with chronic tonsillitis or with hyperplasia reveals disease-associated protein expression differences

  • Original Paper
  • Published:
Analytical and Bioanalytical Chemistry Aims and scope Submit manuscript

Abstract

A proteomic approach has been used to establish a proteome map and differentiate between the protein composition of tonsils from patients with chronic tonsillitis (CT) and that of tonsils with hyperplasia (HPL). Two-dimensional gel analysis was performed with material from four patients with HPL and five patients with CT. An average of approximately 600 spots were detected in each gel. A total of 127 different proteins were identified in 158 spots analyzed by mass spectrometry. Our study revealed disease-associated differences between protein abundance for two protein spots, an HSP27 isoform and UMP-CMP kinase. Both protein spots were more abundant in the CT group. HSP27 ELISA was performed for 32 patients, 12 belonging to the HPL group and 20 to the CT group. ELISA could not be used to differentiate HSP27 isoforms nor to distinguish CT from HPL. HSP27 was found to migrate to two further protein spots in the 2D gels. The differently expressed HSP27 isoform migrated as the most acidic of all the HSP27 isoforms detected, indicating the highest degree of phosphorylation. The sum of all three HSP27 abundances in the gels from the CT group was not different from that of the HPL group, consistent with the ELISA results. Our results suggest that phosphorylation differences caused the observed migration differences of HSP27. Together with the UMP-CMP kinase abundance differences, we conclude that kinase and/or phosphatase activity are different in CT and HPL.

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

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

2DE:

Two-dimensional gel electrophoresis

aa:

Amino acid

ASLO:

Antistreptolysin titer O

CHCA:

α-Cyano-4-hydroxycinnamic acid

CT:

Chronic tonsillitis

CRP:

C-reactive protein

HSP27:

Heat-shock protein 27

HPL:

Hyperplasia

MALDI:

Matrix-assisted laser desorption/ionization

m/z :

Mass-to-charge ratio

SDS-PAGE:

Sodium dodecylsulfate polyacrylamide gel electrophoresis

TFA:

Trifluoroacetic acid

TMB:

Tetramethylbenzidine

TOF-MS:

Time-of-flight mass spectrometry

References

  1. Zhang PC, Pang YT, Loh KS, Wang DY (2003) Clin Otolaryngol 28:235–239

    Article  PubMed  CAS  Google Scholar 

  2. Agren K, Andersson U, Nordlander B, Nord CE, Linde A, Ernberg I, Andersson J (1995) Acta Otolaryngol 115:689–696

    Article  PubMed  CAS  Google Scholar 

  3. Ogino S, Notake N, Harada T, Matsunaga T (1988) Acta Otolaryngol Suppl 454:299–304

    Article  PubMed  CAS  Google Scholar 

  4. Harabuchi Y, Wakashima J, Murakata H, Yoshioka I, Yokoyama Y, Kataura A (1996) Acta Otolaryngol Suppl 523:75–77

    PubMed  CAS  Google Scholar 

  5. Tanaka S, Nakagawa S, Furukawa M (1996) Acta Otolaryngol Suppl 523:78–79

    PubMed  CAS  Google Scholar 

  6. Lorenz P, Bantscheff M, Ibrahim SM, Thiesen HJ, Glocker MO (2003) Clin Chem Lab Med 41:1622–1632

    Article  PubMed  CAS  Google Scholar 

  7. Neuhoff V, Arold N, Taube D, Ehrhardt W (1988) Electrophoresis 9:255–262

    Article  PubMed  CAS  Google Scholar 

  8. Sinz A, Bantscheff M, Mikkat S, Ringel B, Drynda S, Kekow J, Thiesen HJ, Glocker MO (2002) Electrophoresis 23:3445–3456

    Article  PubMed  CAS  Google Scholar 

  9. Fountoulakis M, Langen H (1997) Anal Biochem 250:153–156

    Article  PubMed  CAS  Google Scholar 

  10. Nordhoff E, Egelhofer V, Giavalisco P, Eickhoff H, Horn M, Przewieslik T, Theiss D, Schneider U, Lehrach H, Gobom J (2001) Electrophoresis 22:2844–2855

    Article  PubMed  CAS  Google Scholar 

  11. Nordhoff E, Schürenberg M, Thiele G, Lübbert C, Kloeppel KD, Theiss D, Lehrach H, Gobom J (2003) Intern J Mass Spectrometry 226:163–180

    Article  CAS  Google Scholar 

  12. Garcia M, Platet N, Liaudet E, Laurent V, Derocq D, Brouillet JP, Rochefort H (1996) Stem Cells 14:642–650

    Article  PubMed  CAS  Google Scholar 

  13. Butt E, Immler D, Meyer HE, Kotlyarov A, Laass K, Gaestel M (2001) J Biol Chem 276:7108–7113

    Article  PubMed  CAS  Google Scholar 

  14. Tremolada L, Magni F, Valsecchi C, Sarto C, Mocarelli P, Perego R, Cordani N, Favini P, Galli Kienle M, Sanchez JC, Hochstrasser DF, Corthals GL (2005) Proteomics 5:788–795

    Article  PubMed  CAS  Google Scholar 

  15. Anderson NL, Polanski M, Pieper R, Gatlin T, Tirumalai RS, Conrads TP, Veenstra TD, Adkins JN, Pounds JG, Fagan R, Lobley A (2004) Mol Cell Proteomics 3:311–326

    Article  PubMed  CAS  Google Scholar 

  16. Yui S, Nakatani Y, Mikami M (2003) Biol Pharm Bull 26:753–760

    Article  PubMed  CAS  Google Scholar 

  17. Seeliger S, Vogl T, Engels IH, Schroder JM, Sorg C, Sunderkotter C, Roth J (2003) Am J Pathol 163:947–956

    PubMed  CAS  Google Scholar 

  18. Roth J, Sunderkotter C, Goebeler M, Gutwald J, Sorg C (1992) Int Arch Allergy Immunol 98:140–145

    Article  PubMed  CAS  Google Scholar 

  19. Roth J, Teigelkamp S, Wilke M, Grun L, Tummler B, Sorg C (1992) Immunobiology 186:304–314

    PubMed  CAS  Google Scholar 

  20. Goebeler M, Roth J, Burwinkel F, Vollmer E, Bocker W, Sorg C (1994) Transplantation 58:355–361

    PubMed  CAS  Google Scholar 

  21. Frosch M, Strey A, Vogl T, Wulffraat NM, Kuis W, Sunderkotter C, Harms E, Sorg C, Roth J (2000) Arthritis Rheum 43:628–637

    Article  PubMed  CAS  Google Scholar 

  22. Roth J, Goebeler M, Sorg C (2001) Lancet 357:1041

    Article  PubMed  CAS  Google Scholar 

  23. Roth J, Vogl T, Sorg C, Sunderkotter C (2003) Trends Immunol 24:155–158

    Article  PubMed  CAS  Google Scholar 

  24. Youssef P, Roth J, Frosch M, Costello P, Fitzgerald O, Sorg C, Bresnihan B (1999) J Rheumatol 26:2523–2528

    PubMed  CAS  Google Scholar 

  25. Sugino Y, Teraoka H, Shimono H (1966) J Biol Chem 241:961–969

    PubMed  CAS  Google Scholar 

  26. Liou JY, Dutschman GE, Lam W, Jiang Z, Cheng YC (2002) Cancer Res 62:1624–1631

    PubMed  CAS  Google Scholar 

  27. Van Rompay AR, Johansson M, Karlsson A (1999) Mol Pharmacol 56:562–569

    PubMed  Google Scholar 

  28. Benndorf R, Hayess K, Ryazantsev S, Wieske M, Behlke J, Lutsch G (1994) J Biol Chem 269:20780–20784

    PubMed  CAS  Google Scholar 

  29. Mehlen P, Mehlen A, Guillet D, Preville X, Arrigo AP (1995) J Cell Biochem 58:248–259

    Article  PubMed  CAS  Google Scholar 

  30. Landry J, Lambert H, Zhou M, Lavoie JN, Hickey E, Weber LA, Anderson CW (1992) J Biol Chem 267:794–803

    PubMed  CAS  Google Scholar 

  31. Hickey E, Brandon SE, Potter R, Stein G, Stein J, Weber LA (1986) Nucleic Acids Res 14:4127–4145

    Article  PubMed  CAS  Google Scholar 

  32. Cairns J, Qin S, Philp R, Tan YH, Guy GR (1994) J Biol Chem 269:9176–9183

    PubMed  CAS  Google Scholar 

Download references

Acknowledgements

This work was supported by grants from the University of Rostock and the “Fonds der Chemischen Industrie” to MOG. The authors express their gratitude to Professor Dr H. Nizze of the Institute for Pathology from the University of Rostock. M. Sieb is acknowledged for excellent technical assistance and Mr M. Kreutzer for valuable help with image analysis and quantitative evaluation.

Author information

Author notes

  1. Evariste Gafumbegete

    Present address: Institute and Outpatient Department for Pathology, Ludmillenstift Hospital, Ludmillenstraβe 4–6, 49716, Meppen, Germany

Authors and Affiliations

  1. Department of Otorhinolaryngology, Head and Neck Surgery, Medical Faculty, University of Rostock, 18055, Rostock, Germany

    Tino Just, Jan Gramberg & Hans Wilhelm Pau

  2. Institute for Pathology, Medical Faculty, University of Rostock, 18055, Rostock, Germany

    Evariste Gafumbegete

  3. Proteome Center Rostock, Medical and Natural Science Faculty, University of Rostock, Schillingallee 69,, 18057, Rostock, Germany

    Jan Gramberg, Ines Prüfer, Stefan Mikkat, Bruno Ringel & Michael O. Glocker

Authors
  1. Tino Just
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Evariste Gafumbegete
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Jan Gramberg
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ines Prüfer
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Stefan Mikkat
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Bruno Ringel
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Hans Wilhelm Pau
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Michael O. Glocker
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Michael O. Glocker.

Additional information

This paper was presented at the 38th Annual Meeting of the German Society for Mass Spectrometry (DGMS) held in March 2005 in Rostock, Germany.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Just, T., Gafumbegete, E., Gramberg, J. et al. Differential proteome analysis of tonsils from children with chronic tonsillitis or with hyperplasia reveals disease-associated protein expression differences. Anal Bioanal Chem 384, 1134–1144 (2006). https://doi.org/10.1007/s00216-005-0288-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00216-005-0288-y

Keywords

Search

Navigation