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Proteomes 2014, 2(3), 382-398; doi:10.3390/proteomes2030382

Quantitative Proteomic Profiling of Low-Dose Ionizing Radiation Effects in a Human Skin Model

1
Seattle Genomics, Seattle, WA 98021, USA
2
Environmental Molecular Sciences Laboratory, Pacific Northwest National Laboratory, Richland, WA 99352, USA
3
Fundamental and Computational Sciences Division, Pacific Northwest National Laboratory, Richland, WA 99352, USA
*
Author to whom correspondence should be addressed.
Received: 16 April 2014 / Revised: 8 July 2014 / Accepted: 18 July 2014 / Published: 29 July 2014
(This article belongs to the Special Issue Radiation Proteomics)
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Abstract

To assess responses to low-dose ionizing radiation (LD-IR) exposures potentially encountered during medical diagnostic procedures, nuclear accidents or terrorist acts, a quantitative proteomic approach was used to identify changes in protein abundance in a reconstituted human skin tissue model treated with 0.1 Gy of ionizing radiation. To improve the dynamic range of the assay, subcellular fractionation was employed to remove highly abundant structural proteins and to provide insight into radiation-induced alterations in protein localization. Relative peptide quantification across cellular fractions, control and irradiated samples was performing using 8-plex iTRAQ labeling followed by online two-dimensional nano-scale liquid chromatography and high resolution MS/MS analysis. A total of 107 proteins were detected with statistically significant radiation-induced change in abundance (>1.5 fold) and/or subcellular localization compared to controls. The top biological pathways identified using bioinformatics include organ development, anatomical structure formation and the regulation of actin cytoskeleton. From the proteomic data, a change in proteolytic processing and subcellular localization of the skin barrier protein, filaggrin, was identified, and the results were confirmed by western blotting. This data indicate post-transcriptional regulation of protein abundance, localization and proteolytic processing playing an important role in regulating radiation response in human tissues. View Full-Text
Keywords: ionizing radiation; iTRAQ; quantitative; online 2D LC; filaggrin; skin tissue ionizing radiation; iTRAQ; quantitative; online 2D LC; filaggrin; skin tissue
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MDPI and ACS Style

Hengel, S.M.; Aldrich, J.T.; Waters, K.M.; Pasa-Tolic, L.; Stenoien, D.L. Quantitative Proteomic Profiling of Low-Dose Ionizing Radiation Effects in a Human Skin Model. Proteomes 2014, 2, 382-398.

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