Original Article

Journal of Investigative Dermatology (1994) 103, 364–369; doi:10.1111/1523-1747.ep12394957

Translocation and Downregulation of Protein Kinase C Isoenzymes-alpha and -alt epsilon by Phorbol Ester and Bryostatin-1 in Human Keratinocytes and Fibroblasts

Nicholas J Reynolds1, Joseph J Baldassare2, Patricia A Henderson2, John L Shuler1, Lawrence M Ballas3, David J Burns3, Cindy R Moomaw3 and Gary J Fisher3

  1. 1Departments of Dermatology, University of Michigan, Ann Arbor, Michigan
  2. 2Department of Internal Medicine, St. Louis University, St. Louis, Missouri
  3. 3Sphinx Pharmaceuticals, Durham, North Carolina

Received 12 August 1993; Accepted 8 April 1994.

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Abstract

Protein kinase C isoenzymes can be subdivided into two classes, based on their requirement for calcium. Protein kinase C-alpha, -betaI, -beta, and -gamma are calcium dependent, whereas protein kinase C-gamma, -alt epsilon, -zeta, -eta, and -theta are calcium independent. We have examined the expression, translocation, downregulation, and activation of calcium-dependent and -independent protein kinase C isoenzymes in human skin keratinocytes and fibroblasts. Human keratinocytes and fibroblasts expressed protein kinase C-alpha, -delta, -eta, and -zeta mRNA and protein, whereas protein kinase C-eta (L) was detected only in keratinocytes. Protein kinase C-&, -betaII, -gamma, and -theta were not detected in either cell type. The protein kinase C activators 12-0-tetradecanoylphorbol 13-acetate and bryostatin-1 (50 nM, for 5 min) induced translocation of protein kinase C-alpha and -alt epsilon cytosol to membrane in both keratinocytes and fibroblasts. 12-0-tetradecanoylphorbol 13-acetate and bryostatin-1, for 18 h, induced complete downregulation (i.e., loss) of protein kinase C-alpha and -alt epsilon in keratinocytes, but only partial downregulation was observed in fibroblasts. The subcellular distribution of protein kinase C-alpha, -(or protein kinase C-eta, in keratinocytes or fibroblasts, did not change in response to 12-0-tetradecanoylphorbol 13-acetate or bryostatin-1. These data indicate differential expression, subcellular distribution, and regulation of protein kinase C isoenzymes in human skin cells.

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