title: Using a kinase-inducible bimolecular switch to control enzyme activity in living cells.
abstract: Molecular switches have been instrumental in the development of powerful and versatile genetic tools for directly probing biochemical processes, such as intracellular signaling, within their native contexts. A molecular switch can be broadly defined as a molecular system capable of existing in either of two states (e.g., conformations), which can be converted from one state to the other by a specific input stimulus. This protocol outlines a method for using a kinase-inducible bimolecular switch, along with live-cell fluorescence microscopy, to directly control and monitor the activity of a specific enzyme in living cells.
authors.affiliation: Department of Pharmacology and Molecular Sciences, The Johns Hopkins University School of Medicine, Baltimore, Maryland.
authors.forename: Sohum
authors.initials: S
authors.lastname: Mehta
authors.forename: Jin
authors.initials: J
authors.lastname: Zhang
mesh_terms: D002460:Cell Line; D002468:Cell Physiological Phenomena / Q000235:genetics*; D004247:DNA / Q000235:genetics; D005812:Genes, Switch / Q000235:genetics*; D006801:Humans; D007202:Indicators and Reagents; D008856:Microscopy, Fluorescence; D010770:Phosphotransferases / Q000235:genetics*; D015398:Signal Transduction
chemical_list: D007202:Indicators and Reagents; D004247:DNA; D010770:Phosphotransferases
keywords: biochemical perturbation; biosensor; fluorescence; live-cell imaging