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Protein kinase C in rat cerebral microvessels

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Molecular and Chemical Neuropathology

Abstract

Activation of protein kinase C is a key event in the transduction of receptor-mediated extracellular signals. Little is known about the role of protein kinase C in the microcirculation of the brain. In this study, we examined protein kinase C in isolated cerebral microvessels. A technique for partial purification of protein kinase C from microvessels was employed, using Q-Sepharose batch adsorption and singlestep salt elution in microfuge tubes. This procedure greatly reduced variability and increased protein kinase C specific activity in both the cytosolic and particulate fractions by nearly 50-fold. The identity of the enzyme was confirmed by its inhibition by staurosporine and bisindolylmaleimide and by its translocation in response to phorbol ester. The level of protein kinase C was assessed by [3H]phorbol ester binding and the endogenous substrates evaluated by in vitro phosphorylation studies. Finally, western blot analysis of protein kinase C isoforms indicated that the β-isoform was present in both cytosolic and particulate fractions. The α-isoform was present at low levels in the cytosolic fraction, whereas the ψ-isoform was not detected.

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Authors and Affiliations

  1. Department of Pathology, University of Oklahoma Health Sciences Center, BMSB 434, P.O. Box 26901, 73190, Oklahoma City, OK

    Olivia Hanson-Painton, Kurt Morgenstern, Bobby Moore, Toya Botchlet & Paula Grammas

  2. Department of Chemistry, University of Central Oklahoma, Edmond, OK

    Olivia Hanson-Painton

  3. Department of Medicine, Veterans Hospital, Tampa, FL

    Denise R. Cooper

Authors
  1. Olivia Hanson-Painton
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  2. Kurt Morgenstern
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  3. Denise R. Cooper
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  4. Bobby Moore
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  5. Toya Botchlet
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  6. Paula Grammas
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Hanson-Painton, O., Morgenstern, K., Cooper, D.R. et al. Protein kinase C in rat cerebral microvessels. Molecular and Chemical Neuropathology 20, 245–261 (1993). https://doi.org/10.1007/BF03160077

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  • DOI: https://doi.org/10.1007/BF03160077

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