Inactivation of calcineurin by hydrogen peroxide and phenylarsine oxide. Evidence for a dithiol-disulfide equilibrium and implications for redox regulation - PubMed (original) (raw)

Inactivation of calcineurin by hydrogen peroxide and phenylarsine oxide. Evidence for a dithiol-disulfide equilibrium and implications for redox regulation

R Bogumil et al. Eur J Biochem. 2000 Mar.

Free article

Abstract

Calcineurin (CaN) is a Ca2+-and calmodulin (CaM)-dependent serine/threonine phosphatase containing a dinuclear Fe-Zn center in the active site. Recent studies have indicated that CaN is a possible candidate for redox regulation. The inactivation of bovine brain CaN and of the catalytic CaN A-subunit from Dictyostelium by the vicinal dithiol reagents phenylarsine oxide (PAO) and melarsen oxide (MEL) and by H2O2 was investigated. PAO and MEL inhibited CaN with an IC50 of 3-8 microM and the inactivation was reversed by 2, 3-dimercapto-1-propane sulfonic acid. The treatment of isolated CaN with hydrogen peroxide resulted in a concentration-dependent inactivation. Analysis of the free thiol content performed on the H2O2 inactivated enzyme demonstrated that only two or three of the 14 Cys residues in CaN are modified. The inactivation of CaN by H2O2 could be reversed with 1,4-dithiothreitol and with the dithiol oxidoreductase thioredoxin. We propose that a bridging of two closely spaced Cys residues in the catalytic CaN A-subunit by PAO/MEL or the oxidative formation of a disulfide bridge by H2O2 involving the same Cys residues causes the inactivation. Our data implicate a possible involvement of thioredoxin in the redox control of CaN activity under physiological conditions. The low temperature EPR spectrum of the native enzyme was consistent with a Fe3+-Zn2+ dinuclear centre. Upon H2O2-mediated inactivation of the enzyme no significant changes in the EPR spectrum were observed ruling out that Fe2+ is present in the active enzyme and that the dinuclear metal centre is the target for the oxidative inactivation of CaN.

PubMed Disclaimer

Cited by

Activation of brain calcineurin (Cn) by Cu-Zn superoxide dismutase (SOD1) depends on direct SOD1-Cn protein interactions occurring in vitro and in vivo.
Agbas A, Hui D, Wang X, Tek V, Zaidi A, Michaelis EK. Agbas A, et al. Biochem J. 2007 Jul 1;405(1):51-9. doi: 10.1042/BJ20061202. Biochem J. 2007. PMID: 17324120 Free PMC article.
Defining cutaneous molecular pathobiology of arsenicals using phenylarsine oxide as a prototype.
Srivastava RK, Li C, Weng Z, Agarwal A, Elmets CA, Afaq F, Athar M. Srivastava RK, et al. Sci Rep. 2016 Oct 11;6:34865. doi: 10.1038/srep34865. Sci Rep. 2016. PMID: 27725709 Free PMC article.
Effect of redox balance alterations on cellular localization of LAT and downstream T-cell receptor signaling pathways.
Gringhuis SI, Papendrecht-van der Voort EA, Leow A, Nivine Levarht EW, Breedveld FC, Verweij CL. Gringhuis SI, et al. Mol Cell Biol. 2002 Jan;22(2):400-11. doi: 10.1128/MCB.22.2.400-411.2002. Mol Cell Biol. 2002. PMID: 11756537 Free PMC article.
Thioredoxin 1-mediated post-translational modifications: reduction, transnitrosylation, denitrosylation, and related proteomics methodologies.
Wu C, Parrott AM, Fu C, Liu T, Marino SM, Gladyshev VN, Jain MR, Baykal AT, Li Q, Oka S, Sadoshima J, Beuve A, Simmons WJ, Li H. Wu C, et al. Antioxid Redox Signal. 2011 Nov 1;15(9):2565-604. doi: 10.1089/ars.2010.3831. Epub 2011 Jun 8. Antioxid Redox Signal. 2011. PMID: 21453190 Free PMC article. Review.
Tyrosine Dephosphorylation of ASC Modulates the Activation of the NLRP3 and AIM2 Inflammasomes.
Mambwe B, Neo K, Javanmard Khameneh H, Leong KWK, Colantuoni M, Vacca M, Muimo R, Mortellaro A. Mambwe B, et al. Front Immunol. 2019 Jul 5;10:1556. doi: 10.3389/fimmu.2019.01556. eCollection 2019. Front Immunol. 2019. PMID: 31333677 Free PMC article.

Inactivation of calcineurin by hydrogen peroxide and phenylarsine oxide. Evidence for a dithiol-disulfide equilibrium and implications for redox regulation - PubMed (original) (raw)