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Investigation of Domain Responsible for Calcineurin B Homologous Protein (CHP) Isoform Specific Function
Author(s) -
Davis Shane,
Wallert Mark,
Provost Joseph
Publication year - 2018
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.2018.32.1_supplement.804.22
Subject(s) - gene isoform , microbiology and biotechnology , function (biology) , biology , chemistry , biochemistry , gene
Mammalian cells ubiquitously express the Sodium‐Hydrogen Exchanger (NHE1) a membrane transporter responsible for intracellular pH i , motility and proliferation. Calcineurin B Homologous Protein (CHP) regulates activation and kinetics of the NHE1. CHP has two isoforms, CHP1 and CHP2, whose physiological function remains unclear but both have a unique role in nascent tumor survival. There is a signature domain in the sequence of an otherwise homologous structure between CHP1 and CHP2 that we believe is responsible for CHP isoform specific function, including regulation of NHE1. The purpose of this study is to understand how the these domains of CHP1 and CHP2 confer to the properties of the CHP, particularly cell proliferation and migration. To identify possible differences of structural conformation that could be related to function of each CHP isoform we exchanged the (aa‐aa ofr CHP1 and aa‐aa of CHP2 using gibson cloning. Gibson cloning allows us to manipulate fragments created by PCR to make mutants of our wild‐types. These constructs were then transiently transformed into cells and changes in proliferation and migration analyzed compared to wild‐type expressing cells. We hypothesize that there is an interaction between CHP1 and CHP2 that allow for their individual absence or presence to affect the proliferation and motility function in lung carcinoma. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .