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.
Park
Yong-Doo
YD
Department of Dermatology, Sungkyunkwan University School of Medicine, Samsung Medical Center, Seoul, Korea.
Jang
Hee-Sun
HS
Kim
So-Yeon
SY
Ko
Sung-Kyun
SK
Lyou
You-Jeong
YJ
Lee
Dong-Youn
DY
Paik
Young-Ki
YK
Yang
Jun-Mo
JM
D000293:Adolescent; D000328:Adult; D015415:Biomarkers / Q000378:metabolism; D015153:Blotting, Western; D002478:Cells, Cultured; D002853:Chromatography, Liquid; D003876:Dermatitis, Atopic / Q000378:metabolism*; D015180:Electrophoresis, Gel, Two-Dimensional; D005347:Fibroblasts; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008297:Male; D020543:Proteome / Q000032:analysis*; D040901:Proteomics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012867:Skin / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D015415:Biomarkers; D020543:Proteome