The second hydrophobic domain contributes to the kinetic properties of epithelial sodium channels

Abstract

The epithelial sodium channel (ENaC) is the prototype of a new class of ion channels known as the ENaC/Deg family. The hallmarks of ENaC are a high selectivity for Na+, block by amiloride, small conductance, and slow kinetics that are voltage-independent. We have investigated the contribution of the second hydrophobic domain of each of the homologous subunits α, β, and γ to the kinetic properties of ENaC. Chimeric subunits were constructed between α and β subunits (α-β) and between and β subunits (γ-β). Chimeric and wild-type subunits were expressed in various combinations in Xenopus oocytes. Analysis of whole-cell and unitary currents made it possible to correlate functional properties with specific sequences in the subunits. Functional channels were generated without the second transmembrane domain from α subunits, indicating that it is not essential to form functional pores. The open probability and kinetics varied with the different channels and were influenced by the second hydrophobic domains. Amiloride affinity, Li+/Na+ selectivity, and single channel conductance were also affected by this segment.