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Monitoring calcium‐deficient calmodulin in mouse embryonic stem cells before and after cellular differentiation
Author(s) -
Israel Odisho Koma,
Jervis Eric,
Guillemette Joseph Guy
Publication year - 2009
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.23.1_supplement.683.3
Subject(s) - microbiology and biotechnology , cytoplasm , embryonic stem cell , calmodulin , calcium , calcium binding protein , cell , chemistry , confocal , nucleus , calcium signaling , confocal microscopy , endogeny , cellular differentiation , green fluorescent protein , biology , signal transduction , biochemistry , gene , geometry , mathematics , organic chemistry
Calmodulin (CaM) is a calcium‐sensing protein that binds and regulates over 300 cellular proteins. Cellular localization and trafficking of calcium‐free CaM prior and after Ca 2+ stimulation is not well understood. It has been reported that 95% of total endogenous CaM is bound to target proteins in the cytoplasm, and that the influx of Ca 2+ results in its translocation to the nucleus. The present study was designed to investigate the Ca 2+ ‐ dependent and Ca 2+ ‐independent protein trafficking of CaM during cellular differentiation of mouse embryonic stem cell (MESC). Several CaM mutants were generated and labeled with a fluorescent marker for our investigation. In addition, a FITC labeled cell penetrating peptide (CPP) was used as a tool to introduce these CaM mutants into the cytoplasm of MESCs while maintaining high cell viability. The effects of these modifications on protein trafficking in MESCs during cellular differentiation are monitored via live fluorescent imaging and confocal microscopy. This research is supported by NSERC (J.G.G)