PKC-dependent regulation of transepithelial resistance: roles of MLC and MLC kinase

Am J Physiol. 1999 Sep;277(3):C554-62. doi: 10.1152/ajpcell.1999.277.3.C554.


The mechanisms by which protein kinase C (PKC) activation results in increased transepithelial resistance (TER) are unknown [G. Hecht, B. Robinson, and A. Koutsouris. Am. J. Physiol. 266 (Gastrointest. Liver Physiol. 29): G214-G221, 1994]. We have previously shown that phosphorylation of the regulatory light chain of myosin II (MLC) is associated with decreases in TER and have suggested that contraction of the perijunctional actomyosin ring (PAMR) increases tight junction (TJ) permeability [J. R. Turner, B. K. Rill, S. L. Carlson, D. Carnes, R. Kerner, R. J. Mrsny, and J. L. Madara. Am. J. Physiol. 273 (Cell Physiol. 42): C1378-C1385, 1997]. We therefore hypothesized that PKC activation alters TER via relaxation of the PAMR. Activation of PKC by the phorbol ester phorbol 12-myristate 13-acetate (PMA) resulted in a progressive dose-dependent increase in TER that was apparent within 15 min (111% of controls) and maximal within 2 h (142% of controls). Similar increases were induced by a diacylglycerol analog, and the effects of both PMA and the diacylglycerol analog were prevented by the PKC inhibitor bisindolylmaleimide I. PMA treatment caused progressive decreases in MLC phosphorylation, by 12% at 15 min and 41% at 2 h. Phosphorylation of MLC kinase (MLCK) increased by 64% within 15 min of PMA treatment and was stable over 2 h (51% greater than that of controls). Thus increases in MLCK phosphorylation preceded decreases in MLC phosphorylation. These data suggest that PKC regulates TER via decreased phosphorylation of MLC, possibly due to inhibitory phosphorylation of MLCK. The decreased phosphorylation of MLC likely reduces PAMR tension, leading to decreased TJ permeability.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Biological Transport / drug effects
  • Caco-2 Cells
  • Diglycerides / pharmacology
  • Dose-Response Relationship, Drug
  • Electric Impedance
  • Enzyme Activation / physiology
  • Enzyme Inhibitors / pharmacology
  • Humans
  • Intestinal Mucosa / physiology*
  • Myosin Light Chains / metabolism
  • Myosin Light Chains / physiology*
  • Myosin-Light-Chain Kinase / physiology*
  • Phosphorylation
  • Protein Kinase C / antagonists & inhibitors
  • Protein Kinase C / metabolism
  • Protein Kinase C / physiology*
  • Tetradecanoylphorbol Acetate / pharmacology


  • Diglycerides
  • Enzyme Inhibitors
  • Myosin Light Chains
  • 1-oleoyl-2-acetylglycerol
  • Protein Kinase C
  • Myosin-Light-Chain Kinase
  • Tetradecanoylphorbol Acetate