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Azimzadeh, O. ; Scherthan, H.* ; Yentrapalli, R. ; Barjaktarovic, Z. ; Ueffing, M.* ; Conrad, M. ; Neff, F. ; Calzada-Wack, J. ; Aubele, M. ; Buske, C.* ; Atkinson, M.J. ; Hauck, S.M.* ; Tapio, S.

Label-free protein profiling of formalin-fixed paraffin-embedded (FFPE) heart tissue reveals immediate mitochondrial impairment after ionising radiation.

J. Proteomics 75, 2384-2395 (2012)
DOI
Open Access Green as soon as Postprint is submitted to ZB.
Qualitative proteome profiling of formalin-fixed, paraffin-embedded (FFPE) tissue is advancing the field of clinical proteomics. However, quantitative proteome analysis of FFPE tissue is hampered by the lack of an efficient labelling method. The usage of conventional protein labelling on FFPE tissue has turned out to be inefficient. Classical labelling targets lysine residues that are blocked by the formalin treatment. The aim of this study was to establish a quantitative proteomics analysis of FFPE tissue by combining the label-free approach with optimised protein extraction and separation conditions. As a model system we used FFPE heart tissue of control and exposed C57BL/6 mice after total body irradiation using a gamma ray dose of 3 gray. We identified 32 deregulated proteins (p≤0.05) in irradiated hearts 24h after the exposure. The proteomics data were further evaluated and validated by bioinformatics and immunoblotting investigation. In good agreement with our previous results using fresh-frozen tissue, the analysis indicated radiation-induced alterations in three main biological pathways: respiratory chain, lipid metabolism and pyruvate metabolism. The label-free approach enables the quantitative measurement of radiation-induced alterations in FFPE tissue and facilitates retrospective biomarker identification using clinical archives.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Label-free; Ionising radiation; Proteomics; Mitochondria; FFPE; BOX-BINDING PROTEIN-1; INDUCED GENOMIC INSTABILITY; MASS-SPECTROMETRY; PROTEOMIC ANALYSIS; PRESSURE-OVERLOAD; OXIDATIVE STRESS; EXPRESSION; MEMBRANE; IDENTIFICATION; DYSFUNCTION
ISSN (print) / ISBN 1874-3919
e-ISSN 1876-7737
Quellenangaben Volume: 75, Issue: 8, Pages: 2384-2395 Article Number: , Supplement: ,
Publisher Elsevier
Reviewing status Peer reviewed