Reversible oxidation of phosphatase and tensin homolog (PTEN) alters its interactions with signaling and regulatory proteins
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Published version
Author(s)
Verrastro, I
Tveen-Jensen, K
Woscholski, R
Spickett, CM
Pitt, AR
Type
Journal Article
Abstract
Phosphatase and tensin homolog (PTEN) is involved in a number of different cellular processes including metabolism, apoptosis, cell proliferation and survival. It is a redox-sensitive dual-specificity protein phosphatase that acts as a tumor suppressor by negatively regulating the PI3K/Akt pathway. While direct evidence of redox regulation of PTEN downstream signaling has been reported, the effect of PTEN redox status on its protein–protein interactions is poorly understood. PTEN-GST in its reduced and a DTT-reversible H2O2-oxidized form was immobilized on a glutathione-sepharose support and incubated with cell lysate to capture interacting proteins. Captured proteins were analyzed by LC–MSMS and comparatively quantified using label-free methods. 97 Potential protein interactors were identified, including a significant number that are novel. The abundance of fourteen interactors was found to vary significantly with the redox status of PTEN. Altered binding to PTEN was confirmed by affinity pull-down and Western blotting for Prdx1, Trx, and Anxa2, while DDB1 was validated as a novel interactor with unaltered binding. These results suggest that the redox status of PTEN causes a functional variation in the PTEN interactome. The resin capture method developed had distinct advantages in that the redox status of PTEN could be directly controlled and measured.
Date Issued
2016-01-01
Date Acceptance
2015-11-05
Citation
Free Radical Biology and Medicine, 2016, 90, pp.24-34
ISSN
0891-5849
Publisher
Elsevier
Start Page
24
End Page
34
Journal / Book Title
Free Radical Biology and Medicine
Volume
90
Copyright Statement
© 2015 The Authors. Published by Elsevier Inc. This is an open access article under the CC BY license
(http://creativecommons.org/licenses/by/4.0/)
(http://creativecommons.org/licenses/by/4.0/)
Subjects
Science & Technology
Life Sciences & Biomedicine
Biochemistry & Molecular Biology
Endocrinology & Metabolism
Protein oxidation
Mass spectrometry
Proteomics
Protein-protein interactions
Peroxiredoxin-1
Redox biology
Signaling
Thioredoxin-1
TUMOR-SUPPRESSOR PTEN
SPECTROMETRY-BASED PROTEOMICS
FATTY-ACID SYNTHASE
MASS-SPECTROMETRY
HEPATOCELLULAR-CARCINOMA
SIZE CHECKPOINT
PROSTATE-CANCER
IN-VITRO
CELLS
PATHWAY
Publication Status
Published
Date Publish Online
2015-11-10