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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)
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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Publikationstyp
Artikel: Journalartikel
Dokumenttyp
Wissenschaftlicher Artikel
Schlagwörter
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
Zeitschrift
Journal of Proteomics
Quellenangaben
Band: 75,
Heft: 8,
Seiten: 2384-2395
Verlag
Elsevier
Nichtpatentliteratur
Publikationen
Begutachtungsstatus
Peer reviewed