Second transmembrane domain modulates epithelial sodium channel gating in response to shear stress

Abi-Antoun T, Shi S, Tolino LA, Kleyman TR, Carattino MD. Second transmembrane domain modulates epithelial sodium channel gating in response to shear stress. Am J Physiol Renal Physiol 300: F1089-F1095, 2011. First published February 9, 2011; doi:10.1152/ajprenal.00610.2010.-Na+ absorption and K+ secretion in the distal segments of the nephron are modulated by the tubular flow rate. Epithelial Na+ channels (ENaC), composed of alpha-, beta-, and gamma-subunits respond to laminar shear stress (LSS) with an increase in open probability. Higher vertebrates express a delta-ENaC subunit that is functionally related to the alpha-subunit, while sharing only 35% of sequence identity. We investigated the response of delta beta gamma channels to LSS. Both the time course and magnitude of activation of delta beta gamma channels by LSS were remarkably different from those of alpha beta gamma channels. ENaC subunits have similar topology, with an extracellular region connected by two transmembrane domains with intracellular N and C termini. To identify the specific domains that are responsible for the differences in the response to flow of alpha beta gamma and delta beta gamma channels, we generated a series of alpha-delta chimeras and site-specific alpha-subunit mutants and examined parameters of activation by LSS. We found that specific sites in the region encompassing and just preceding the second transmembrane domain were responsible for the differences in the magnitude and time course of channel activation by LSS.