Article thumbnail

α, ß–unsaturated carbonyls covalently modify cellular PTEN.

By Tracy M. Covey (171796), Kornelia Edes (360526), Gary S. Coombs (360527), David M. Virshup (171835) and Frank A. Fitzpatrick (360528)

Abstract

<p>(<b>A</b>) Diagram of the procedure to identify PTEN with an oxidized or alkylated thiol in cells exposed to ROS or α, ß–unsaturated carbonyls. The anti-PTEN immunoblot shows oxidized or carbonylated PTEN (NA Pulldown) relative to total PTEN (Cell Lysate) isolated from MCF-7 cells treated 30 min with vehicle (DMSO), 10 µM Δ12-PGJ<sub>2</sub> or 4-HNE versus 10 min with 100 µM H<sub>2</sub>O<sub>2</sub>; an immunoblot from a separate experiment shows oxidized, carbonylated and total PTEN in cells treated for 30 min with vehicle or 20 µM acrolein versus 10 min with 100 µM H<sub>2</sub>O<sub>2</sub>. (<b>B</b>) Anti-PTEN immunoblot of MCF-7 cell lysates fractionated by SDS-PAGE under non-reducing conditions. PTEN oxidized to a Cys<sup>124</sup>-Cys<sup>71</sup> disulfide appears as a faster migrating species (PTEN oxidized disulfide) only in cells treated with H<sub>2</sub>O<sub>2</sub>. This species was undetectable in MCF-7cells treated 30 min with DMSO vehicle or 20 µM each Δ12-PGJ<sub>2</sub>, 4-HNE, acrolein or 15-HpETE. (<b>C</b>) Chemical structures of typical α, ß–unsaturated carbonyls. Acrolein and 4-HNE are α, β enals; Δ12PGJ<sub>2</sub> is an α,β enone. Electrophilic β carbons are denoted with δ<sup>+</sup>. Blots are representative of results obtained in at least three independent experiments.</p

Topics: Biochemistry, Cell Biology, Immunology, Cancer, carbonyls, covalently, cellular
Year: 2013
DOI identifier: 10.1371/journal.pone.0013545.g001
OAI identifier: oai:figshare.com:article/493091
Provided by: FigShare
Download PDF:
Sorry, we are unable to provide the full text but you may find it at the following location(s):
  • https://figshare.com/articles/... (external link)
  • Suggested articles


    To submit an update or takedown request for this paper, please submit an Update/Correction/Removal Request.