Effects of H2O2 on protein tyrosine phosphatase activity in HER14 cells

Sullivan, Stephen Gene; Chiu, Daniel T.; Errasfa, Mourad; Wang, Jamin M.; Qi, Jian-Shen; Stern, Arnold

doi:10.1016/0891-5849(94)90042-6

Cited by 114 publications

(55 citation statements)

References 22 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…The active site cysteines of various PTPs, including Cys 215 of PTP1B, are specifically targeted by the sulfhydryl-modifying reagent iodoacetic acid (12-14, 16, 17). The same active site cysteine residues have also been implicated as the site of oxidation by various oxidants (2,3,6,7,29), in which case one should be able to monitor the extent of H 2 O 2 -induced inactivation of PTPs in cells by measuring the amount of radioactivity incorporated into the enzyme after cell lysis in the presence of radiolabeled iodoacetic acid. With this aim, we expressed the 37-kDa PTP1B in E. coli, purified the recombinant protein, and subjected it to modification by H 2 O 2 and iodoacetic acid.…”

Section: Resultsmentioning

confidence: 99%

“…Because the extent of protein tyrosine phosphorylation in a cell reflects an equilibrium between the actions of protein-tyrosine kinases (PTKs) and protein-tyrosine phosphatases (PTPs), either stimulation of PTKs or inhibition of PTPs would be expected to shift the equilibrium toward phosphorylation. PTP activity in crude cell extracts can be inactivated by various oxidants, including H 2 O 2 , and this inactivation can be reversed by incubation with thiol compounds such as dithiothreitol (DTT) and GSH (6,7). These observations suggest that PTPs might undergo H 2 O 2 -dependent inactivation in cells, resulting in a shift in the equilibrium with PTKs toward phosphorylation.…”

mentioning

confidence: 94%

See 1 more Smart Citation

Reversible Inactivation of Protein-tyrosine Phosphatase 1B in A431 Cells Stimulated with Epidermal Growth Factor

Lee¹,

Kwon²,

Kim³

et al. 1998

Journal of Biological Chemistry

901

671

View full text Add to dashboard Cite

Stimulation of various cells with growth factors results in a transient increase in the intracellular concentration of H 2 O 2 that is required for growth factor-induced protein tyrosine phosphorylation. The effect of H 2 O 2 produced in response to epidermal growth factor (EGF) on the activity of protein-tyrosine phosphatase 1B (PTP1B) was investigated in A431 human epidermoid carcinoma cells. H 2 O 2 inactivated recombinant PTP1B in vitro by oxidizing its catalytic site cysteine, most likely to sulfenic acid. The oxidized enzyme was reactivated more effectively by thioredoxin than by glutaredoxin or glutathione at their physiological concentrations. Oxidation by H 2 O 2 prevented modification of the catalytic cysteine of PTP1B by iodoacetic acid, suggesting that it should be possible to monitor the oxidation state of PTP1B in cells by measuring the incorporation of radioactivity into the enzyme after lysis of the cells in the presence of radiolabeled iodoacetic acid. The amount of such radioactivity associated with PTP1B immunoprecipitated from A431 cells that had been stimulated with EGF for 10 min was 27% less than that associated with PTP1B from unstimulated cells. The amount of iodoacetic acid-derived radioactivity associated with PTP1B reached a minimum 10 min after stimulation of cells with EGF and returned to base line values by 40 min, suggesting that the oxidation of PTP1B is reversible in cells. These results indicate that the activation of a receptor tyrosine kinase by binding of the corresponding growth factor may not be sufficient to increase the steady state level of protein tyrosine phosphorylation in cells and that concurrent inhibition of protein-tyrosine phosphatases by H 2 O 2 might also be required.

show abstract

Section: Resultsmentioning

confidence: 99%

mentioning

confidence: 94%

Reversible Inactivation of Protein-tyrosine Phosphatase 1B in A431 Cells Stimulated with Epidermal Growth Factor

Lee¹,

Kwon²,

Kim³

et al. 1998

Journal of Biological Chemistry

901

671

View full text Add to dashboard Cite

show abstract

“…This dose range of H 2 O 2 used in cancerous or transformed cells is considered to be appropriate in mimicking oxidative stress conditions [28,31,[57][58][59] . Bossis et al reported that ROS at low concentrations, 1 mmol/L, result in the rapid disappearance of most SUMO conjugates, which is due to direct and reversible inhibition of SUMO conjugating enzymes through the formation of (a) disulfide bond(s) involving the catalytic cysteines of the SUMO E1 subunit Uba2 and the E2-conjugating enzyme Ubc9.…”

Section: Discussionmentioning

confidence: 99%

The biphasic redox sensing of SENP3 accounts for the HIF-1 transcriptional activity shift by oxidative stress

Wang

Yang

et al. 2012

Acta Pharmacol Sin

View full text Add to dashboard Cite

“…It is likely that the effects of exposing cells to H 2 O 2 are mediated by global inhibition of tyrosine phosphatases (27)(28)(29)(30)(31) 7 Jurkat cells (10 6 cells/ml) to a final concentration of 5 mM. After incubation at 37°C for 15 min, the cells were washed once in ice-cold, isotonic, Tris-buffered saline and then lysed in RIPA buffer (150 mM NaCl, 50 mM Tris-HCl, pH 7.2, 1% w/v sodium deoxycholate, 1% Nonidet P-40, 0.1% SDS, 200 M Na 3 VO 4 , 50 mM NaF, 2 mM EDTA, 100 kallikrein-inactivating units/ml aprotinin) for 20 min at 4°C at a concentration of 10 7 cells/ml.…”

Section: From the ‡Molecular Biology And Virology Laboratory The Salmentioning

confidence: 99%

The Activated Form of the Lck Tyrosine Protein Kinase in Cells Exposed to Hydrogen Peroxide Is Phosphorylated at Both Tyr-394 and Tyr-505

Hardwick

Sefton

1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

Members of the Src family of non-receptor tyrosine protein kinases are known to be inhibited by the intramolecular association between a phosphorylated carboxyl-terminal tyrosine residue and the SH2 domain. We have previously shown that exposure of cells to H 2 O 2 strongly activates Lck, a lymphocyte-specific Src family kinase, by inducing phosphorylation on Tyr-394, an absolutely conserved residue within the activation loop of the catalytic domain. Here we show that Lck that has been activated by H 2 O 2 is simultaneously phosphorylated at both the carboxyl-terminal tyrosine (Tyr-505) and Tyr-394. Thus, dephosphorylation of Tyr-505 is not a prerequisite for either phosphorylation of Lck at Tyr-394 or catalytic activation of the kinase. These results indicate that activation of Lck by phosphorylation of Tyr-394 is dominant over any inhibition induced by phosphorylation of Tyr-505. We propose that these results may be extended to all Src family members.p56 lck , a member of the Src family of non-receptor tyrosine protein kinases (1, 2), is expressed predominantly in T cells. Lck function is critical both for T-cell development in the thymus (3, 4) and activation of mature T cells in the periphery by antigen (5, 6). Lck stably associates with the inner surface of the plasma membrane as a result of myristoylation of Gly-2 and palmitoylation of Ser-3 and Ser-5 (7-10). There it binds to the T-cell receptor-associated glycoproteins CD4 and CD8 as well as other proteins through its unique amino terminus (11-15). The activity of Lck is regulated by phosphorylation of two conserved tyrosine residues. Tyr-505 (equivalent to Tyr-527 in c-Src) is located near the carboxyl terminus of Lck and, when phosphorylated, associates intramolecularly with the SH2 domain in the amino-terminal half of the protein. This helps stabilize Lck in a conformation that, biologically, is relatively inactive (16 -20). In the absence of phosphorylation at Tyr-505, intramolecular binding of the carboxyl terminus to the SH2 domain does not occur, and Lck exhibits increased activity in vivo. In contrast, phosphorylation of Tyr-394 (equivalent to Tyr-416 in c-Src) stimulates the catalytic activity of Lck (21-23). Phosphorylation of Tyr-394 allows the formation of hydrogen bonds between the phosphate of Tyr(P) 1 -394 and amino acid residues in the catalytic cleft. These interactions allow the enzyme to assume a conformation resembling that of activated cyclic AMP-dependent protein kinase A (19, 20, 24 -26).We have previously demonstrated that hydrogen peroxide, a potent activator of Lck, acts by inducing phosphorylation of Lck on 37). It is likely that the effects of exposing cells to H 2 O 2 are mediated by global inhibition of tyrosine phosphatases (27-31). The increase in phosphorylation of Lck at Tyr-394 that we observe in the presence of H 2 O 2 may therefore result largely from reduced dephosphorylation of this site. Our previous work did not address the question of whether or not activation of Lck by H 2 O 2 -induced phosphorylation of Tyr-394...

show abstract

Effects of H2O2 on protein tyrosine phosphatase activity in HER14 cells

Cited by 114 publications

References 22 publications

Reversible Inactivation of Protein-tyrosine Phosphatase 1B in A431 Cells Stimulated with Epidermal Growth Factor

Reversible Inactivation of Protein-tyrosine Phosphatase 1B in A431 Cells Stimulated with Epidermal Growth Factor

The biphasic redox sensing of SENP3 accounts for the HIF-1 transcriptional activity shift by oxidative stress

The Activated Form of the Lck Tyrosine Protein Kinase in Cells Exposed to Hydrogen Peroxide Is Phosphorylated at Both Tyr-394 and Tyr-505

Contact Info

Product

Resources

About