Protein kinase C isoforms in human aortic smooth muscle cells

J. J. Grange, L. M. Baca-Regen, A. J. Nollendorfs, Y. Persidsky, D. L. Sudan, B. T. Baxter, J. Alexander

Research output: Contribution to journalArticlepeer-review

9 Scopus citations

Abstract

Purpose: To identify the protein kinase C (PKC) isoforms in human arterial smooth muscle cells (SMC) and define their subcellular location in the resting state and in response to the PKC activator, 12-O- tetradecanoylphorbol 13-acetate (TPA). Methods: Arterial SMC cultures established from transplant donor aorta were treated with 100 nM TPA or control media, then mechanically lysed. PKC from the soluble and particulate fraction were separated by centrifugation, and protein normalized immunoblots were performed with antibodies to the PKC isoforms α, β(I), β(II), δ, ε, γ and ζ. Bands were detected by enhanced chemiluminescence and analyzed densitometrically, with results expressed as the mean percentage of each fraction ± SEM. Translocation was defined as a significant (p < 0.05) change in the particulate fraction for each isoform. Immunofluorescent staining of cultured SMC visualized the resting location and stimulated translocation of each isoform. Results: Isoforms α and β1 were detected primarily in the soluble fraction, translocating to the particulate fraction with TPA stimulation (p < 0.0001). The isoforms β(II), δ, and ε were found primarily in the particulate fraction and did not translocate. Immunofluorescent staining confirmed these locations. Neither γ or ζ were detected in these SMC. Conclusions: The PKC isoforms expressed in human arterial SMC differ from those reported in animal models. Their specific locations and response to stimulation suggest unique functions in cellular regulation and provide the groundwork for further investigation into their role in the development of vascular disease and regulation of matrix metabolism.

Original languageEnglish (US)
Pages (from-to)919-927
Number of pages9
JournalJournal of vascular surgery
Volume27
Issue number5
DOIs
StatePublished - 1998

ASJC Scopus subject areas

  • Surgery
  • Cardiology and Cardiovascular Medicine

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