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Am J Physiol Cell Physiol 275: C848-C856, 1998;
0363-6143/98 $5.00
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Vol. 275, Issue 3, C848-C856, September 1998

Characterization of the cloned human intermediate-conductance Ca2+-activated K+ channel

Bo Skaaning Jensen1, Dorte Strøbæk1, Palle Christophersen1, Tino Dyhring Jørgensen1, Claus Hansen2, Asli Silahtaroglu2,3, Søren-Peter Olesen1, and Philip Kiær Ahring1

1 NeuroSearch A/S, DK-2600 Glostrup; 2 Department of Medical Genetics, Institute of Medical Biochemistry and Genetics, University of Copenhagen, DK-2200 Copenhagen N, Denmark; and 3 Department of Genetics, Division of Biomedical Sciences, Cerrahpasa Medical Faculty, Istanbul University, TR-34303 Istanbul, Turkey

The human intermediate-conductance, Ca2+-activated K+ channel (hIK) was identified by searching the expressed sequence tag database. hIK was found to be identical to two recently cloned K+ channels, hSK4 and hIK1. RNA dot blot analysis showed a widespread tissue expression, with the highest levels in salivary gland, placenta, trachea, and lung. With use of fluorescent in situ hybridization and radiation hybrid mapping, hIK mapped to chromosome 19q13.2 in the same region as the disease Diamond-Blackfan anemia. Stable expression of hIK in HEK-293 cells revealed single Ca2+-activated K+ channels exhibiting weak inward rectification (30 and 11 pS at -100 and +100 mV, respectively). Whole cell recordings showed a noninactivating, inwardly rectifying K+ conductance. Ionic selectivity estimated from bi-ionic reversal potentials gave the permeability (PK/PX) sequence K+ = Rb+ (1.0) > Cs+ (10.4) >>  Na+, Li+, N-methyl-D-glucamine (>51). NH+4 blocked the channel completely. hIK was blocked by the classical inhibitors of the Gardos channel charybdotoxin (IC50 28 nM) and clotrimazole (IC50 153 nM) as well as by nitrendipine (IC50 27 nM), Stichodactyla toxin (IC50 291 nM), margatoxin (IC50 459 nM), miconazole (IC50 785 nM), econazole (IC50 2.4 µM), and cetiedil (IC50 79 µM). Finally, 1-ethyl-2-benzimidazolinone, an opener of the T84 cell IK channel, activated hIK with an EC50 of 74 µM.

intermediate-conductance calcium-activated potassium channel; charybdotoxin; clotrimazole; fluorescent in situ hybridization; radiation hybrid mapping; patch clamp; Diamond-Blackfan anemia


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