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Identification of essential residues in the extracellular region of the epithelial sodium channel (997.4)
Author(s) -
Berman Chance,
Booth Rachell
Publication year - 2014
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.28.1_supplement.997.4
Subject(s) - epithelial sodium channel , mutant , extracellular , pseudohypoaldosteronism , renal sodium reabsorption , reabsorption , chemistry , intracellular , yeast , sodium , extracellular fluid , mutation , microbiology and biotechnology , biology , biochemistry , gene , organic chemistry
The epithelial sodium channel (ENaC) is a membrane bound protein responsible for regulating the reabsorption of sodium in the distal nephron of the kidneys. Genetic mutations in the intracellular regions of ENaC are known to cause diseases such as Liddle’s syndrome and Pseudohypoaldosteronism type 1, severe hypertension or hypotension, respectively. In an effort to identify critical residues in the extracellular region of αENaC that are necessary for proper function, error prone polymerase chain reaction (EP‐PCR) was employed to generate random mutations in the extracellular loop followed by expression in yeast. A serial dilution pronging assay was used to monitor the salt sensitivity, an induced phenotype, in yeast expressing various αENaCs. Mutant αENaCs that altered yeast growth were sequenced and expression was confirmed by western blotting. Mutations effecting αENaC function included mutant 125, Y334H; mutant 135, F306L; and mutant 137, R202A. Future studies are aimed at understanding the role of these residues in ENaC function. Grant Funding Source : Supported by NIH R15 GM86798 (R. Booth & W. David) and Welch Foundation Departmental Grant AI‐0045