Thioredoxin reductase 1 and NADPH directly protect protein tyrosine phosphatase 1B from inactivation during H2O2 exposure

Markus Dagnell, Paul E. Pace, Qing Cheng, Jeroen Frijhoff, Arne Ostman, Elias S. J. Arner, Mark B. Hampton, Christine C. Winterbourn*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

Abstract

Regulation of growth factor signaling involves reversible inactivation of protein tyrosine phosphatases (PTPs) through the oxidation and reduction of their active site cysteine. However, there is limited mechanistic understanding of these redox events and their co-ordination in the presence of cellular antioxidant networks. Here we investigated interactions between PTP1B and the peroxiredoxin 2 (Prx2)/thioredoxin 1 (Trx1)/thioredoxin reductase 1 (TrxR1) network. We found that Prx2 becomes oxidized in PDGF-treated fibroblasts, but only when TrxR1 has first been inhibited. Using purified proteins, we also found that PTP1B is relatively insensitive to inactivation by H2O2 but found no evidence for a relay mechanism in which Prx2 or Trx1 facilitates PTP1B oxidation. Instead, these proteins prevented PTP1B inactivation by H2O2. Intriguingly, we discovered that TrxR1/NADPH directly protects PTP1B from inactivation when present during the H2O2 exposure. This protection was dependent on the concentration of TrxR1 and independent of Trx1 and Prx2. The protection was blocked by auranofin and required an intact selenocysteine residue in TrxR1. This activity likely involves reduction of the sulfenic acid intermediate form of PTP1B by TrxR1 and is therefore distinct from the previously described reactivation of end-point oxidized PTP1B, which requires both Trx1 and TrxR1. The ability of TrxR1 to directly reduce an oxidized phosphatase is a novel activity that can help explain previously observed increases in PTP1B oxidation and PDGF receptor phosphorylation in TrxR1 knockout cells. The activity of TrxR1 is therefore of potential relevance for understanding the mechanisms of redox regulation of growth factor signaling pathways.

Original languageEnglish
Pages (from-to)14371-14380
Number of pages10
JournalJournal of Biological Chemistry
Volume292
Issue number35
DOIs
Publication statusPublished - 1 Sept 2017

Keywords

  • GROWTH-FACTOR
  • HYDROGEN-PEROXIDE
  • REACTIVE OXYGEN
  • REVERSIBLE INACTIVATION
  • SIGNAL-TRANSDUCTION
  • OXIDATIVE STRESS
  • REDOX REGULATION
  • OXIDIZED PTP1B
  • PEROXIREDOXIN
  • SYSTEM

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