title: Rapid Detection of Dynamic PTEN Regulation in Living Cells Using Intramolecular BRET.
abstract: Tumor suppressor PTEN phosphatase acts to inhibit the PI3K/AKT pathway and thus regulates cell proliferation, survival, and migration. Dysregulation of PTEN function is observed in a wide range of cancers. In addition to alterations of the PTEN gene, repression of PTEN function can also occur at the protein level through changes in PTEN conformation, localization, activity, and stability. The ability to follow switches in PTEN conformation in live cells provides a rapid approach to study changes in PTEN function and may provide a basis to screen pharmacological agents aimed at enhancing or reestablishing PTEN-dependent signaling pathways that have gone awry in cancer. Here, we describe methods to use an intramolecular bioluminescent resonance energy transfer (BRET)-based biosensor that reports dynamic signal-dependent changes in PTEN conformational rearrangement and function.
authors.affiliation: Inserm, U1016, Institut Cochin, 27, Rue du Faubourg Saint Jacques, 75014, Paris, France.|CNRS, UMR8104, Paris, France.|University Paris Descartes, Sorbonne Paris Cité, Paris, France.
authors.forename: Stanislas
authors.initials: S
authors.lastname: Misticone
authors.affiliation: Inserm, U1016, Institut Cochin, 27, Rue du Faubourg Saint Jacques, 75014, Paris, France.|CNRS, UMR8104, Paris, France.|University Paris Descartes, Sorbonne Paris Cité, Paris, France.|Structural Genomics Consortium, University of Toronto, Toronto, ON, Canada.
authors.forename: Evelyne
authors.initials: E
authors.lastname: Lima-Fernandes
authors.affiliation: Inserm, U1016, Institut Cochin, 27, Rue du Faubourg Saint Jacques, 75014, Paris, France. mark.scott@inserm.fr.|CNRS, UMR8104, Paris, France. mark.scott@inserm.fr.|University Paris Descartes, Sorbonne Paris Cité, Paris, France. mark.scott@inserm.fr.
authors.forename: Mark G H
authors.initials: MG
authors.lastname: Scott
mesh_terms: D060807:Bioluminescence Resonance Energy Transfer Techniques / Q000379:methods*; D045744:Cell Line, Tumor; D057809:HEK293 Cells; D006367:HeLa Cells; D006801:Humans; D051059:PTEN Phosphohydrolase / Q000737:chemistry*
chemical_list: D051059:PTEN Phosphohydrolase; C494929:PTEN protein, human
keywords: Bioluminescence resonance energy transfer; Biosensor; Conformational change; PI3K/AKT; PTEN; Phosphatase; Posttranslational modifications; Tumor suppressor