Abstract
PTEN is a one of the most frequently mutated tumor suppressors in human cancers. It is essential for regulating diverse biological processes and through its lipid phosphatase activity regulates the PI 3-Kinase signaling pathway. Sensitive phosphatase assays are employed to study the catalytic activity of PTEN against phospholipid substrates. Here we describe protocols to assay PTEN lipid phosphatase activity using either purified enzyme (purified PTEN lipid phosphatase assay) or PTEN immunopurified from tissues or cultured cells (cellular IP PTEN lipid phosphatase assay) against vesicles containing radiolabeled PIP3 substrate.
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Acknowledgements
We would like to thank the members of the NRL laboratory for their constructive discussions. Work developing and applying these assays was funded by the Medical Research Council (grant code G0801865).
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Spinelli, L., Leslie, N.R. (2016). Assays to Measure PTEN Lipid Phosphatase Activity In Vitro from Purified Enzyme or Immunoprecipitates. In: Pulido, R. (eds) Protein Tyrosine Phosphatases. Methods in Molecular Biology, vol 1447. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3746-2_6
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DOI: https://doi.org/10.1007/978-1-4939-3746-2_6
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Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3744-8
Online ISBN: 978-1-4939-3746-2
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