Proteomic analysis of the response of EpiDerm cultures to sodium lauryl sulphate.
The analysis of EpiDerm cultures treated with the known skin irritant sodium lauryl sulphate (SLS) was performed using 2D-gel electrophoresis in order to understand the mechanism of action and thereby identify novel markers of skin irritation. A range of both broad and narrow pH gradient first-dimension gels were run (pH 4-7, 6-11, 4-5, 5-6 and 6-9) consistently followed by 12% SDS-PAGE in the second-dimension. Following treatment of EpiDerm with SLS, 67 proteins of interest were identified, of which 8 were selected as interesting: calmodulin-like skin protein, involucrin, epithelial cell marker protein, HS1, peroxiredoxin 1, serine protease inhibitor, KIAA0117 and ribosomal protein L17. Involucrin was confirmed as being up-regulated by both ELISA and Western blotting. The use of proteomics has identified a number of proteins which could be used as general markers for skin irritation and which may in particular be of value for the development of in vitro predictive models.
Fletcher
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SEAC-Safety and Environmental Assurance Centre, Unilever Colworth, Sharnbrook, Bedfordshire MK44 1LQ, UK. Samantha.Fletcher@Unilever.com
Basketter
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D015153:Blotting, Western; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional; D004797:Enzyme-Linked Immunosorbent Assay; D055551:HSP27 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000096:biosynthesis; D006801:Humans; D007509:Irritants / Q000633:toxicity*; D015603:Keratinocytes / Q000187:drug effects*; D018832:Molecular Chaperones; D009363:Neoplasm Proteins / Q000096:biosynthesis; D040901:Proteomics; D015842:Serine Proteinase Inhibitors / Q000096:biosynthesis; D012967:Sodium Dodecyl Sulfate / Q000633:toxicity*
D055551:HSP27 Heat-Shock Proteins; C057725:HSPB1 protein, human; D006360:Heat-Shock Proteins; D007509:Irritants; D018832:Molecular Chaperones; D009363:Neoplasm Proteins; D015842:Serine Proteinase Inhibitors; D012967:Sodium Dodecyl Sulfate