Suppression of Kir2.3 activity by protein kinase C phosphorylation of the channel protein at threonine 53

J Biol Chem. 1999 Apr 23;274(17):11643-6. doi: 10.1074/jbc.274.17.11643.


Kir2.3 plays an important part in the maintenance of membrane potential in neurons and myocardium. Identification of intracellular signaling molecules controlling this channel thus may lead to an understanding of the regulation of membrane excitability. To determine whether Kir2.3 is modulated by direct phosphorylation of its channel protein and identify the phosphorylation site of protein kinase C (PKC), we performed experiments using several recombinant and mutant Kir2.3 channels. Whole-cell Kir2.3 currents were inhibited by phorbol 12-myristate 13-acetate (PMA) in Xenopus oocytes. When the N-terminal region of Kir2.3 was replaced with that of Kir2.1, another member in the Kir2 family that is insensitive to PMA, the chimerical channel lost its PMA sensitivity. However, substitution of the C terminus was ineffective. Four potential PKC phosphorylation sites in the N terminus were studied by comparing mutations of serine or threonine with their counterpart residues in Kir2.1. Whereas substitutions of serine residues at positions 5, 36, and 39 had no effect on the channel sensitivity to PMA, mutation of threonine 53 completely eliminated the channel response to PMA. Interestingly, creation of this threonine residue at the corresponding position (I79T) in Kir2.1 lent the mutant channel a PMA sensitivity almost identical to the wild-type Kir2.3. These results therefore indicate that Kir2.3 is directly modulated by PKC phosphorylation of its channel protein and threonine 53 is the PKC phosphorylation site in Kir2.3.

Publication types

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

MeSH terms

  • Animals
  • Female
  • Mutagenesis, Site-Directed
  • Phosphorylation
  • Potassium Channel Blockers*
  • Potassium Channels / chemistry
  • Potassium Channels / genetics
  • Potassium Channels, Inwardly Rectifying*
  • Protein Kinase C / metabolism*
  • Tetradecanoylphorbol Acetate / pharmacology
  • Threonine / antagonists & inhibitors*
  • Xenopus laevis


  • KCNJ4 protein, human
  • Potassium Channel Blockers
  • Potassium Channels
  • Potassium Channels, Inwardly Rectifying
  • Threonine
  • Protein Kinase C
  • Tetradecanoylphorbol Acetate