Chapter title |
Rapid Detection of Dynamic PTEN Regulation in Living Cells Using Intramolecular BRET
|
---|---|
Chapter number | 8 |
Book title |
PTEN
|
Published in |
Methods in molecular biology, January 2016
|
DOI | 10.1007/978-1-4939-3299-3_8 |
Pubmed ID | |
Book ISBNs |
978-1-4939-3297-9, 978-1-4939-3299-3
|
Authors |
Stanislas Misticone, Evelyne Lima-Fernandes, Mark G. H. Scott |
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. |
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