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Mutational analysis of calcineurin B homologous protein isoform 2 binding to the Na + ‐ H + (796.8)
Author(s) -
Pantera Harrison,
Bisnett Brittany,
Hasle Daniel,
Cottle Wayne,
Provost Joseph,
Wallert Mark
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.796.8
Subject(s) - gene isoform , microbiology and biotechnology , mutant , chemistry , biology , biochemistry , gene
The Na + ‐H + exchanger isoform 1 (NHE1) is a transmembrane protein that regulates a range of cellular functions essential for cancer progression including cell adhesion, proliferation and migration. The calcineurin B homologous proteins (CHP1 and CHP2) appear to be essential cofactors to support NHE1 function. The CHP1 and CHP 2 binding domain on NHE1 is the same, amino acids 515 to 530. CHP2 is expressed primarily in tumor cells where it binds to NHE1 with a 5‐10 fold higher affinity than CHP1. CHP2 expression in tumor cells supports increased invasion and migration. Here we investigate the ability of mutations to a key amino acid in the binding domain (N519) to alter CHP2 binding to NHE1 in vivo . Two distinct site directed mutations to NHE1, N519A and N519D, have been constructed along with the accompanying stable cell lines expressing NHE1 with these mutations. We will present data evaluating CHP2 binding to NHE1 in these cells using a GFP‐CHP construct. We will also evaluate how a change in CHP2 binding alters adhesion, proliferation, and migration in mutant expressing cells as compared to cells expressing wild‐type NHE1.