Two-dimensional electrophoretic profiling of atopic dermatitis in primary cultured keratinocytes from patients

Recently, we reported altered protein expression in primary cultured fibroblasts from atopic dermatitis (AD) patients. As a sequential study, we conducted proteomic analysis of primary keratinocytes derived from AD patients to further identify AD‐related proteins. Three pH ranges, 4–7, 6–9, and 7–11, were used to profile the altered protein expression in AD. We obtained 46 candidate spots from the 2‐D gel image analysis: 18 proteins were up‐regulated and 27 proteins were down‐regulated. Among the several important candidate proteins, NCC27 showed the same profile of a defect in PTM in both AD‐derived keratinocytes and fibroblasts. On the basis of current and previous reports, real‐time PCR was performed on select candidate genes to compare RNA and protein expression levels in AD‐derived keratinocytes and fibroblasts. Our results provide new clues to aid in understanding the mechanism of atopic alterations in keratinocytes and suggest new AD‐associated proteins that are important in AD pathogenesis.