Protein kinase Cα modulates depolarizaton-evoked changes of intracellular Ca2+ concentration in a rat pheochromocytoma cell line

A. M. Fontainhas, A. G. Obukhov, Martha Nowycky

Research output: Contribution to journalArticlepeer-review

8 Scopus citations


Conventional protein kinase C (cPKC) isoforms are activated by a coincident rise in cytosolic Ca2+ and membrane-bound diacylglycerol. In excitable cells, cPKC may be activated by Ca2= influx through voltage-gated Ca2+ channels (VGCC). cPKCs, in turn, are known to modulate the activity of VGCC. We examined whether PKCα, a cPKC, could be activated by depolarization in a neuroendocrine cell line and whether activation occurred on a time scale that modulated the depolarization-evoked intracellular Ca2+ concentration ([Ca2+]i) signal. Pheochromocytoma cells (PC12 cells) were transfected with wild-type and mutant forms of PKCα labeled with yellow fluorescent protein to monitor kinase translocation. Simultaneously, [Ca2+]i changes were monitored with fura-2. Two point mutations that render PKCα inactive, D187A in the Ca2+ binding site and K368R in the ATP binding site, significantly prolonged the time-to-peak of the depolarization-evoked [Ca 2+]i signal. A mutation that modulates membrane insertion (W58G) and two mutations of an autophosphorylation site (S657A, S657E) had no effect on the kinetics of the [Ca2+]i signal. We conclude that in PC12 cells, Ca2+ entry through VGCC rapidly activates PKCα, and that PKCα can modulate the Ca2+ signal on a physiologically relevant time scale. Point mutations of PKCα can be used as specific and potent modulators of the PKC signaling pathway.

Original languageEnglish (US)
Pages (from-to)393-403
Number of pages11
Issue number2
StatePublished - Jun 7 2005

All Science Journal Classification (ASJC) codes

  • Neuroscience(all)


  • Depolarization
  • Fura-2
  • PC12 cells
  • Voltage-gated Ca channels
  • cPKC

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