title: Quantitative phosphoproteomics identifies filaggrin and other targets of ionizing radiation in a human skin model.
abstract: Our objective here was to perform a quantitative phosphoproteomic study on a reconstituted human skin tissue to identify low- and high-dose ionizing radiation-dependent signalling in a complex three-dimensional setting. Application of an isobaric labelling strategy using sham and three radiation doses (3, 10, 200 cGy) resulted in the identification of 1052 unique phosphopeptides. Statistical analyses identified 176 phosphopeptides showing significant changes in response to radiation and radiation dose. Proteins responsible for maintaining skin structural integrity including keratins and desmosomal proteins (desmoglein, desmoplakin, plakophilin 1, 2 and 3) had altered phosphorylation levels following exposure to both low and high doses of radiation. Altered phosphorylation of multiple sites in profilaggrin linker domains coincided with altered profilaggrin processing suggesting a role for linker phosphorylation in human profilaggrin regulation. These studies demonstrate that the reconstituted human skin system undergoes a coordinated response to both low and high doses of ionizing radiation involving multiple layers of the stratified epithelium that serve to maintain tissue integrity and mitigate effects of radiation exposure.
authors.affiliation: Pacific Northwest National Laboratory, Richland, WA 99352, USA.
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
authors.forename: Katrina M
authors.initials: KM
authors.lastname: Waters
authors.forename: Bobbie-Jo
authors.initials: BJ
authors.lastname: Webb-Robertson
authors.forename: Marianne B
authors.initials: MB
authors.lastname: Sowa
authors.forename: Claere
authors.initials: C
authors.lastname: von Neubeck
authors.forename: Josh T
authors.initials: JT
authors.lastname: Aldrich
authors.forename: Lye Meng
authors.initials: LM
authors.lastname: Markillie
authors.forename: Rachel M
authors.initials: RM
authors.lastname: Wirgau
authors.forename: Marina A
authors.initials: MA
authors.lastname: Gritsenko
authors.forename: Rui
authors.initials: R
authors.lastname: Zhao
authors.forename: David G
authors.initials: DG
authors.lastname: Camp
authors.forename: Richard D
authors.initials: RD
authors.lastname: Smith
authors.forename: David L
authors.initials: DL
authors.lastname: Stenoien
mesh_terms: D051182:Desmogleins / Q000378:metabolism; D051180:Desmoplakins / Q000378:metabolism; D004307:Dose-Response Relationship, Radiation; D000091344:Filaggrin Proteins; D006801:Humans; D007381:Intermediate Filament Proteins / Q000378:metabolism*; D007633:Keratins / Q000378:metabolism; D010748:Phosphopeptides / Q000378:metabolism*; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation / Q000528:radiation effects; D051191:Plakophilins / Q000378:metabolism; D040901:Proteomics; D011839:Radiation, Ionizing; D015398:Signal Transduction / Q000528:radiation effects; D012867:Skin / Q000378:metabolism* / Q000528:radiation effects*
chemical_list: D051182:Desmogleins; D051180:Desmoplakins; C000717249:FLG protein, human; D000091344:Filaggrin Proteins; D007381:Intermediate Filament Proteins; D010748:Phosphopeptides; D010750:Phosphoproteins; D051191:Plakophilins; D007633:Keratins