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

  • Tino Just
  • Evariste Gafumbegete
  • Jan Gramberg
  • Ines Prüfer
  • Stefan Mikkat
  • Bruno Ringel
  • Hans Wilhelm Pau
  • Michael O. GlockerEmail author
Original Paper

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.

Keywords

Tonsil hyperplasia Recurrent/chronic tonsillitis Proteomics Mass spectrometry MALDI-MS HSP27 Phosphorylation 

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

Notes

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.

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  • Tino Just
    • 1
  • Evariste Gafumbegete
    • 2
  • Jan Gramberg
    • 1
    • 3
  • Ines Prüfer
    • 3
  • Stefan Mikkat
    • 3
  • Bruno Ringel
    • 3
  • Hans Wilhelm Pau
    • 1
  • Michael O. Glocker
    • 3
    Email author
  1. 1.Department of Otorhinolaryngology, Head and Neck Surgery, Medical FacultyUniversity of RostockRostockGermany
  2. 2.Institute for Pathology, Medical FacultyUniversity of RostockRostockGermany
  3. 3.Proteome Center Rostock, Medical and Natural Science FacultyUniversity of RostockRostockGermany

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