Redox regulation of type-I inositol trisphosphate receptors in intact mammalian cells.

ABSTRACT: A sensitization of inositol 1,4,5-trisphosphate receptor (IP₃R)-mediated Ca²⁺ release is associated with oxidative stress in multiple cell types. These effects are thought to be mediated by alterations in the redox state of critical thiols in the IP₃R, but this has not been directly demonstrated in intact cells. Here, we utilized a combination of gel-shift assays with MPEG-maleimides and LC-MS/MS to monitor the redox state of recombinant IP₃R1 expressed in HEK293 cells. We found that under basal conditions, ∼5 of the 60 cysteines are oxidized in IP₃R1. Cell treatment with 50 μm thimerosal altered gel shifts, indicating oxidation of ∼20 cysteines. By contrast, the shifts induced by 0.5 mm H₂O₂ or other oxidants wer

SUBMITTER: Joseph SK 

PROVIDER: S-EPMC6231128 | biostudies-literature | 2018 Nov

REPOSITORIES: biostudies-literature