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The connexin43 gap junction protein is phosphorylated by protein kinase A and protein kinase C: In vivo and in vitro studies

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Abstract

There is general agreement that the connexin43 gap junction protein is a substrate for phosphorylation by protein kinase C but there is no similar consensus regarding the action of protein kinase A. Our previous studies demonstrated that channels formed by connexin43 were reversibly gated in response to microinjected protein kinase A and protein kinase C, but we did not determine whether these effects involved direct action on the connexin43 protein. Using a combination of in vivo metabolic labeling and in vitro phosphorylation of recombinant protein and synthetic peptides, we now find that connexin43 is a relatively poor substrate for purified protein kinase A compared to protein kinase C, but that phosphorylation can be accelerated by 8-Br-cAMP (8-bromoadenosine 3′,5′-cyclic monophosphate) which also enhances connexin43 synthesis but at a much slower rate than phosphorylation. Phosphorylation of a critical amino acid, Ser364, by protein kinase A, appears to be necessary for subsequent multiple phosphorylations by protein kinase C. However, protein kinase C can phosphorylate connexin43 at a reduced level in the absence of prior phosphorylation. The results suggest that the correct regulation of channels formed by connexin43 may require sequential phosphorylations of this protein by protein kinase A and protein kinase C.

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Authors and Affiliations

  1. Department of Physiology, Loma Linda University School of Medicine, Division of Anatomy, Loma Linda, CA, 92354, USA

    Maithili M. Shah & William H. Fletcher

  2. Department of Pathology and Human Anatomy, Division of Anatomy, Loma Linda University School of Medicine, Loma Linda, CA, 92354, USA

    Anna-Marie Martinez

  3. Medical Research Service, Jerry L. Pettis Memorial VA Medical Center, Loma Linda, CA, 92357, USA

    Anna-Marie Martinez

Authors
  1. Maithili M. Shah
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  2. Anna-Marie Martinez
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  3. William H. Fletcher
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Shah, M.M., Martinez, AM. & Fletcher, W.H. The connexin43 gap junction protein is phosphorylated by protein kinase A and protein kinase C: In vivo and in vitro studies. Mol Cell Biochem 238, 57–68 (2002). https://doi.org/10.1023/A:1019902920693

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