Artikel
Differential proteome analysis of tonsil material from children with recurrent acute or chronic tonsillitis in comparison with hyperplasia in order to identify diagnostic marker proteins
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Autoren
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Tino Just
- Department of Otorhinolaryngology, Head and Neck Surgery, University of Rostock, Germany
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Evariste Gafumbegete
- Institute for Pathology, University of Rostock
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Jan Gramberg
- Proteome Center Rostock, University of Rostock
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Ines Prüfer
- Proteome Center Rostock, University of Rostock
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Bruno Ringel
- Proteome Center Rostock, University of Rostock
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Stefan Mikkat
- Proteome Center Rostock, University of Rostock
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Hans Wilhelm Pau
- Department of Otorhinolaryngology, Head and Neck Surgery, University of Rostock, Germany
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Michael O. Glocker
- Proteome Center Rostock, University of Rostock
| Veröffentlicht: | 22. September 2005 |
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Gliederung
Text
Differential proteome analysis was used to compare protein pattern from fresh-frozen tonsils derived from children with recurrent acute or chronic tonsillitis, and tonsil hyperplasia, respectively. In order to find disease associated marker proteins, protein pattern of both entities were compared.
Palatine tonsils from 37 children, aged between 2 and 11, were removed surgically. Thirty-seven extracts from those children were analyzed (23 from recurrent acute tonsillitis and 14 from hyperplasia). Two-dimensional gel electrophoresis (2-DE) of the soluble proteins was performed from hyperplasia (n = 2) and recurrent tonsillitis (n = 5) in triplicate. Differential computer-based 2-DE-gel analysis was carried out using two 2D reference gels of each entity. Selected protein spots were excised from the gels, digested with trypsin and identified by peptide mass fingerprinting with MALDI-TOF MS.
600 Protein spots were visualized routinely on each gel using Coomassie staining. A strong similarity in abundance of protein spots within each group was apparent. Only those differences that persisted in the rigorous differential analysis were addressed as differentially expressed, resulting in rather small differences in protein patterns between tonsillitis and hyperplasia. Clear abundance differences between the two groups were found for heat shock protein 27 (HSP27) and UMP-CMP kinase. The spots of both proteins were more intensive in recurrent tonsillitis. HSP27 ELISA analyses were performed to verify the different abundance of HSP27 with extracts from 32 frozen juvenile tonsils. Higher HSP27 level were found in children with tonsillitis in comparison to children with hyperplasia. In further investigations we will evaluate the applicability of HSP27 as diagnostic marker for tonsil disease.
