Using a genetically encoded FRET-based reporter to visualize calcineurin phosphatase activity in living cells.
Calcineurin is an evolutionarily conserved, ubiquitously expressed protein phosphatase that serves as a major effector of Ca(2+) signals, regulating diverse biological processes such as gene expression, tissue differentiation, immune responses, and neural plasticity. The following method describes how to monitor real-time calcineurin activity in cultured mammalian cells using a fluorescence resonance energy transfer (FRET)-based activity reporter.
Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Sohum
S
Mehta
Jin
J
Zhang
D002470:Cell Survival; D004247:DNA / Q000235:genetics; D031541:Fluorescence Resonance Energy Transfer; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D057054:Molecular Imaging / Q000379:methods*; D050778:NFATC Transcription Factors / Q000737:chemistry / Q000235:genetics*; D010744:Phosphoric Monoester Hydrolases / Q000378:metabolism*; D010957:Plasmids / Q000235:genetics; D014162:Transfection
D050778:NFATC Transcription Factors; D004247:DNA; C120817:calcineurin phosphatase; D010744:Phosphoric Monoester Hydrolases