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MBC in Press, published online ahead of print December 22, 2004
Mol. Biol. Cell 10.1091/mbc.E04-06-0537

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Submitted on June 30, 2004
Revised on November 30, 2004
Accepted on December 7, 2004

Identification and Functional Characterization of Ankyrin-Repeat Family Protein, ANKRA, as a Protein Interacting with BKCa Channel

Hyun-Ho Lim and Chul-Seung Park*

Department of Life Science, Gwangju Institute of Science and Technology (GIST), Gwangju 500-712, Korea

Monitoring Editor: Guido Guidotti

Ankyrin-repeat family A protein (ANKRA) was originally cloned in mouse as an interacting protein to megalin, a member of low-density lipoprotein receptor superfamily. Here we report that the isolation of rat ANKRA as a new binding partner for the {alpha}-subunit of rat large-conductance Ca2+-activated K+ channel (rSlo). We mapped the binding region of each protein using yeast two-hybrid and in vitro binding assays. ANKRA expressed together with rSlo channels were colocalized near the plasma membrane and coimmunoprecipitated in transfected cells. We also showed that BKCa channel in rat cerebral cortex coprecipitated with rANKRA and colocalized in cultured rat hippocampal neuron. While the coexpression of ANKRA did not affect the surface expression of rSlo, the gating kinetics of rSlo channel was significantly altered and the effects were highly dependent on the intracellular calcium. These results indicate that ANKRA could modulate the excitability of neurons by binding directly to endogenous BKCa channel and altering its gating kinetics in a calcium dependent manner.

*Corresponding author. E-mail: cspark{at}gist.ac.kr




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