The Effects of Accessory Proteins on ENaC Function
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Maintaining homeostasis is crucial for perpetuating good health. Any imbalance, such as hypertension, can lead to heart and kidney disease. Epithelial sodium channels, also known as ENaC, constitute the rate-limiting step of sodium reabsorption in the distal tubules of the nephron in the kidneys. It is here the final, yet critical, 3% to 5% of sodium reabsorption that dictates blood pressure occurs. In this study, β-ENaC was cloned into pESC-Leu and pESC-Leu/γ in order to further characterize accessory proteins in yeast screens using the heterotrimeric channel. An antibody screen against the subunits of ENaC was then performed using Murine Principle Kidney Cortical Collecting Duct (mpkCCD) cells as a means of identifying a primary antibody for western blotting. RNA Interference studies in mpkCCD cells were also performed. Knockdown of the accessory proteins TMED2 and TMP21 through RNAi indicated a decrease in ENaC expression. These results indicated that TMED2 and TMP21 are essential for ENaC trafficking.