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Molecular and Cellular Biochemistry

, Volume 314, Issue 1–2, pp 161–169 | Cite as

Identification of different isoforms of 14-3-3 protein family in human dermal and epidermal layers

  • Ruhangiz T. Kilani
  • Abelardo Medina
  • Alastair Aitken
  • Reza B. Jalili
  • Matthew Carr
  • Aziz Ghahary
Article

Abstract

We have previously demonstrated a high level of stratifin, also known as 14-3-3 sigma (σ) in differentiated keratinocyte cell lysate and conditioned medium (CM). In this study, we asked the question of whether other 14-3-3 isoforms are expressed in human dermal fibroblasts, keratinocytes, intact dermal and epidermal layers of skin. In order to address this question, total proteins extracted from cultured cells or skin layers were subjected to western blot analysis using seven different primary antibodies specific to well-known mammalian isoforms, β, γ, ε, η, σ, τ, and ζ of 14-3-3 protein family. The autoradiograms corresponding to each isoform were then quantified and compared. The results revealed the presence of very high levels of all seven isoforms in cultured keratinocyte and conditioned medium. With the exception of τ isoform, other 14-3-3 isoforms were also present in intact epidermal layer of normal skin. The profile of 14-3-3 proteins in whole skin was similar to that of epidermis. In contrast, only gamma (γ) 14-3-3 isoform, was present in dermal layer obtained from the same skin sample. On the other hand, cultured fibroblasts express a high level of β, ε, γ and η and a low level of ζ and τ, but not σ isoform. However, the levels of 14-3-3 ε, γ and η were barely detectable in fibroblast conditioned medium. Further, we also used immunohistochemical staining to identify the 14-3-3 isoform expressing cells in human skin sections. The finding revealed different expression profile for each of these isoforms mainly in differentiated keratinocytes located within the layer of lucidum. However, fibroblasts located within the dermal layer did not show any detectable levels of these proteins. In conclusion, all members of 14-3-3 proteins are expressed by cells of epidermal but not dermal layer of skins and that these proteins are mainly expressed by differentiated keratinocytes.

Keywords

Protein 14-3-3s Dermis Epidermis Human skin Keratinocyte Fibroblasts 

Notes

Acknowledgment

This research was supported by the grant MOP-13387 from Canadian Institute for Health Research.

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Copyright information

© Springer Science+Business Media, LLC. 2008

Authors and Affiliations

  • Ruhangiz T. Kilani
    • 1
  • Abelardo Medina
    • 1
  • Alastair Aitken
    • 2
  • Reza B. Jalili
    • 1
  • Matthew Carr
    • 1
  • Aziz Ghahary
    • 1
  1. 1.BC Professional Fire Fighters’ Burn and Wound Healing Laboratory, Department of SurgeryJack Bell Research Centre, University of British ColumbiaVancouverCanada
  2. 2.School of Biological SciencesThe University of EdinburghEdinburghScotland, UK

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