N-acetyltransferase in human skin and keratinocytes

Tanja Vogel, Jutta Bonifas, Marjon Wiegman, Hendrikus Pas, Brunhilde Blömeke, Pieter Jan Coenraads, Marie-Louise Schuttelaar

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Background: N-acetyltransferase 1 (NAT1) mediated Nacetylation in human skin and keratinocytes is an important detoxification pathway for aromatic amines including the strong sensitizer para-phenylenediamine (PPD), an important component of oxidative hair dyes. Objectives: Human skin and keratinocytes have high Nacetylation capacities, however the localization of NAT1 in the epidermis and the influence differentiation on NAT1 is not clear. NAT1 expression and cell proliferation were suggested to be associated in breast and prostate cells, which prompted us to analyze NAT1 in skin and proliferating versus differentiated keratinocytes. Methods: HaCaT and primary keratinocytes (NHEK, five donors) were in vitro differentiated using calcium and differentiation was confirmed by increased Keratin 1, Keratin 10 and Transglutaminase 1 mRNA expression detected by real-time PCR. NAT1 activities were detected in cell lysates and conversion of PPD to its acetylated derivatives was analyzed in cell culture supernatants by HPLC. NAT1 expression in cryosections of human skin obtained from mamma reductions were detected using fluorescent staining with anti-NAT1 primary antibody and fluorescein isothiocyanate goat anti-rabbit IgG secondary antibody. Results: In vitro differentiation of keratinocyte did not alter NAT1 activities, which varied between 21.1±4 and 26.2±3 nmol/mg/min in HaCaT and 7±0.6 and 5.9±0.6 nmol/mg/min in NHEK. Formation of acetylated PPD was demonstrated in supernatants of proliferating as well as differentiated keratinocytes. In keratinocytes an increased conversion of PPD to acetylated derivatives was seen. In healthy human skin staining against NAT1 showed granular staining of the cytosol of the basal layers of keratinocytes. Conclusions: NAT1 expression was demonstrated in human epidermis and overall N-acetylation capacity and consequently detoxification capacitieswas not reduced during in vitro keratinocyte differentiation. NAT1 is expressed in a granular pattern in the cytoplasmic matrix of basal keratinocytes. It could be hypothesized that NAT1 is present in cytosolic vesicles.However, further research on this topic is needed.
Original languageEnglish
Pages (from-to)88-89
Number of pages2
JournalCONTACT DERMATITIS
Volume70
Issue numbers1
DOIs
Publication statusPublished - Jun-2014

Keywords

  • Experimental
  • Others
  • acyltransferase
  • antibody
  • tuberculin
  • phenylenediamine
  • cytokeratin 1
  • cytokeratin 10
  • protein glutamine gamma glutamyltransferase
  • hair dye
  • calcium
  • aromatic amine
  • immunoglobulin G
  • fluorescein isothiocyanate
  • fluorescent dye
  • messenger RNA
  • human
  • skin
  • keratinocyte
  • society
  • contact dermatitis
  • in vitro study
  • detoxification
  • breast
  • epidermis
  • male
  • staining
  • sensitization
  • acetylation
  • donor
  • cytosol
  • prostate
  • normal human
  • rabbit
  • goat
  • cell proliferation
  • basement membrane
  • cell culture
  • real time polymerase chain reaction
  • high performance liquid chromatography

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