Uncovering the Critical Amino Acids in the Beta Bubunit of the Epithelial Sodium Channel (ENaC)
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A high intake of sodium into the body can lead to high blood pressure that could result to stroke, kidney and heart disease. Sodium balance in the body is partially regulated in the distal tubules of the nephron in the kidney via a membrane channel called the epithelial sodium channel (ENaC). The structure, regulation and assembly of ENaC’s subunits is not yet fully understood. In this study, error prone polymerase chain reaction (EP-PCR) was used to introduce random mutations in the extracellular loop of the beta subunit of ENaC in order to identify residues that may be critical to proper function. A yeast screen, dilution survival pronging assay, was used to visualize the level of growth inhibition of yeast cells expressing α-ENaC (i.e. ENaC function) with wild type or various β-ENaC mutants. Also, a western blot was used to confirm mutated ENaC expression in yeast cells. The potential β-ENaC were identified in these preliminary studies and should be further characterized in additional dilution pronging assays and expression studies.