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

被引：31

作者：

Azimzadeh, Omid ^{[1
]}

Scherthan, Harry ^{[2
]}

Yentrapalli, Ramesh ^{[1
,3
]}

Barjaktarovic, Zarko ^{[1
]}

Ueffing, Marius ^{[4
,5
]}

Conrad, Marcus ^{[6
,7
]}

Neff, Frauke ^{[8
]}

Calzada-Wack, Julia ^{[8
]}

Aubele, Michaela ^{[8
]}

Buske, Christian ^{[9
]}

Atkinson, Michael J. ^{[1
,10
]}

Hauck, Stefanie M. ^{[4
]}

Tapio, Soile ^{[1
]}

机构：

[1] German Res Ctr Environm Hlth, Helmholtz Zentrum Munchen, Inst Radiat Biol, D-85764 Neuherberg, Germany

[2] Univ Ulm, Bundeswehr Inst Radiobiol, Munich, Germany

[3] Stockholm Univ, Dept Genet Microbiol & Toxicol, Ctr Radiat Protect Res, S-10691 Stockholm, Sweden

[4] German Res Ctr Environm Hlth, Helmholtz Zentrum Munchen, Res Unit Prot Sci, D-85764 Neuherberg, Germany

[5] Univ Med Ctr, Ctr Ophthalmol, Tubingen, Germany

[6] German Ctr Neurodegenerat Dis, DZNE, Neuherberg, Germany

[7] Helmholtz Zentrum Munchen, Inst Dev Genet, Neuherberg, Germany

[8] German Res Ctr Environm Hlth, Helmholtz Zentrum Munchen, Inst Pathol, D-85764 Neuherberg, Germany

[9] Univ Hosp Ulm, Inst Expt Canc Res, Ulm, Germany

[10] Tech Univ Munich, Klinikum Rechts Isar, Dept Radiat Oncol, D-8000 Munich, Germany

来源：

JOURNAL OF PROTEOMICS | 2012年 / 75卷 / 08期

关键词：

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;

D O I：

10.1016/j.jprot.2012.02.019

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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 24 h 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. (C) 2012 Elsevier B.V. All rights reserved.

引用

页码：2384 / 2395

页数：12

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