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Am J Physiol Lung Cell Mol Physiol. June 15, 2012; 302 (12): L1262-72.

Characterization of a novel splice variant of δ ENaC subunit in human lungs.

Zhao RZ , Nie HG , Su XF , Han DY , Lee A , Huang Y , Chang Y , Matalon S , Ji HL .

Salt absorption via apical epithelial sodium channels (ENaC) is a critical rate-limiting process in maintaining airway and lung lining fluid at the physiological level. δ ENaC (termed δ1 in this article) has been detected in human lung epithelial cells in addition to α, β, and γ subunits (Ji HL, Su XF, Kedar S, Li J, Barbry P, Smith PR, Matalon S, Benos DJ. J Biol Chem 281: 8233-8241, 2006; Nie HG, Chen L, Han DY, Li J, Song WF, Wei SP, Fang XH, Gu X, Matalon S, Ji HL, J Physiol 587: 2663-2676, 2009) and may contribute to the differences in the biophysical properties of amiloride-inhibitable cation channels in pulmonary epithelial cells. Here we cloned a splicing variant of the δ1 ENaC, namely, δ2 ENaC in human bronchoalveolar epithelial cells (16HBEo). δ2 ENaC possesses 66 extra amino acids attached to the distal amino terminal tail of the δ1 ENaC. δ2 ENaC was expressed in both alveolar type I and II cells of human lungs as revealed by in situ hybridization and real-time RT-PCR. To characterize the biophysical and pharmacological features of the splicing variant, we injected Xenopus oocytes with human ENaC cRNAs and measured whole cell and single channel currents of δ1βγ, δ2βγ, and αβγ channels. Oocytes injected with δ2βγ cRNAs exhibited whole cell currents significantly greater than those expressing δ1βγ and αβγ channels. Single channel activity, unitary conductance, and open probability of δ2βγ channels were significantly greater compared with δ1βγ and αβγ channels. In addition, δ2βγ and δ1βγ channels displayed significant differences in apparent Na(+) affinity, dissociation constant for amiloride (K(i)(amil)), the EC(50) for capsazepine activation, and gating kinetics by protons. Channels comprising of this novel splice variant may contribute to the diversities of native epithelial Na(+) channels.

PubMed ID: 22505667
PMC ID: PMC3379047
Article link: Am J Physiol Lung Cell Mol Physiol.
Grant support: ES015676 NIEHS NIH HHS , ES017218 NIEHS NIH HHS , HL031197 NHLBI NIH HHS , HL095435 NHLBI NIH HHS , HL87017 NHLBI NIH HHS , R01 GM085237 NIGMS NIH HHS , ES017218 NIEHS NIH HHS , HL87017 NHLBI NIH HHS , ES015676 NIEHS NIH HHS , HL031197 NHLBI NIH HHS , R01 GM085237 NIGMS NIH HHS , HL095435 NHLBI NIH HHS

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