Protein Kinase C Regulates the Phosphorylation and Cellular Localization of Occludin

Anna Y. Andreeva, Eberhard Krause, Eva Christina Müller, Ingolf E. Blasig, Darkhan I. Utepbergenov

Research output: Contribution to journalArticle

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Abstract

Occludin is an integral membrane phosphoprotein specifically associated with tight junctions, contributing to the structure and function of this intercellular seal. Occludin function is thought to be regulated by phosphorylation, but no information is available on the molecular pathways involved. In the present study, the involvement of the protein kinase C pathway in the regulation of the phosphorylation and cellular distribution of occludin has been investigated. Phorbol 12-myristate 13-acetate and 1,2-dioctanoylglycerol induced the rapid phosphorylation of occludin in Madin-Darby canine kidney cells cultured in low extracellular calcium medium with a concomitant translocation of occludin to the regions of cell-cell contact. The extent of occludin phosphorylation as well as its incorporation into tight junctions induced by protein kinase C activators or calcium switch were markedly decreased by the protein kinase C inhibitor GF-109203X. In addition, in vitro experiments showed that the recombinant COOH-terminal domain of murine occludin could be phosphorylated by purified protein kinase C. Ser338 of occludin was identified as an in vitro protein kinase C phosphorylation site using peptide mass fingerprint analysis and electrospray ionization tandem mass spectroscopy. These findings indicate that protein kinase C is involved in the regulation of occludin function at tight junctions.

Original languageEnglish
Pages (from-to)38480-38486
Number of pages7
JournalJournal of Biological Chemistry
Volume276
Issue number42
DOIs
Publication statusPublished - Oct 19 2001

Fingerprint

Occludin
Phosphorylation
Protein Kinase C
Tight Junctions
Calcium
Tight Junction Proteins
Electrospray ionization
Madin Darby Canine Kidney Cells
Peptide Mapping
Protein C Inhibitor
Phosphoproteins
Protein Kinase Inhibitors
Seals
Mass Spectrometry
Acetates
Switches
Spectroscopy
Membranes
Peptides

ASJC Scopus subject areas

  • Biochemistry

Cite this

Protein Kinase C Regulates the Phosphorylation and Cellular Localization of Occludin. / Andreeva, Anna Y.; Krause, Eberhard; Müller, Eva Christina; Blasig, Ingolf E.; Utepbergenov, Darkhan I.

In: Journal of Biological Chemistry, Vol. 276, No. 42, 19.10.2001, p. 38480-38486.

Research output: Contribution to journalArticle

Andreeva, Anna Y. ; Krause, Eberhard ; Müller, Eva Christina ; Blasig, Ingolf E. ; Utepbergenov, Darkhan I. / Protein Kinase C Regulates the Phosphorylation and Cellular Localization of Occludin. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 42. pp. 38480-38486.
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