Premium
Identification of Extracellular Residues Critical to the Epithelial Sodium Channel
Author(s) -
Dismuke Taylor Y,
Booth Rachell
Publication year - 2017
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.31.1_supplement.772.8
Subject(s) - epithelial sodium channel , reabsorption , extracellular , renal sodium reabsorption , yeast , function (biology) , homeostasis , kidney , sodium , chemistry , microbiology and biotechnology , biology , biochemistry , endocrinology , organic chemistry
Disruption of homeostatic sodium reabsorption takes place in the kidney can result in various diseases such as osteoporosis, kidney stones, or hypertension. The protein channel found in the membrane of kidney cells that carries out sodium reabsorption is the epithelial sodium channel (ENaC). Identifying and characterizing residues that are critical to ENaC function are key to understanding ENaC's mechanism of action. Complete understanding of the structure‐function relationships within ENaC's extracellular loop are still rudimentary, but studies have demonstrated that it plays a role in channel regulation. Using random mutagenesis, mutations within the extracellular loop of βENaC were generated and incorporated into a yeast expression system. Qualitative screening using a survival pronging dilution assay with yeast expressing αβγENaC was used to monitor ENaC function. Yeast cells expressing αβ Mut γENaC demonstrated changes in growth compared to yeast expressing wild‐type αβγENaC. Future studies include sequencing to identify mutations and localization of αβ MUT γENaC within yeast cells. Support or Funding Information This research was supported by National Institute of General Medical Sciences (NIGMS) of the National Institutes of Health under award number R15GM086798 (R. Booth and W. David) and R25GM10783 (K. Oyajobi, N. Blake, R. Booth, and R. Walter).