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Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3

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Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3. / Bassi, Rekha; Burgoyne, Joseph R; DeNicola, Gian F et al.

In: Journal of Biological Chemistry, Vol. 292, No. 39, 29.09.2017, p. 16161-16173.

Research output: Contribution to journalArticlepeer-review

Harvard

Bassi, R, Burgoyne, JR, DeNicola, GF, Rudyk, O, DeSantis, V, Charles, RL, Eaton, P & Marber, MS 2017, 'Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3', Journal of Biological Chemistry, vol. 292, no. 39, pp. 16161-16173. https://doi.org/10.1074/jbc.M117.785410

APA

Bassi, R., Burgoyne, J. R., DeNicola, G. F., Rudyk, O., DeSantis, V., Charles, R. L., Eaton, P., & Marber, M. S. (2017). Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3. Journal of Biological Chemistry, 292(39), 16161-16173. https://doi.org/10.1074/jbc.M117.785410

Vancouver

Bassi R, Burgoyne JR, DeNicola GF, Rudyk O, DeSantis V, Charles RL et al. Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3. Journal of Biological Chemistry. 2017 Sep 29;292(39):16161-16173. https://doi.org/10.1074/jbc.M117.785410

Author

Bassi, Rekha ; Burgoyne, Joseph R ; DeNicola, Gian F et al. / Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3. In: Journal of Biological Chemistry. 2017 ; Vol. 292, No. 39. pp. 16161-16173.

Bibtex Download

@article{9bec5c14896a4f34803b74f75f13bab4,
title = "Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3",
abstract = "The kinase p38α MAPK (p38α) plays a pivotal role in many biological processes. p38α is activated by canonical upstream kinases that phosphorylate the activation region. The purpose of our study was to determine whether such activation may depend on redox-sensing cysteines within p38α. p38α was activated and formed a disulfide-bound heterodimer with MAP2K3 (MKK3) in rat cardiomyocytes and isolated hearts exposed to H2O2 This disulfide heterodimer was sensitive to reduction by mercaptoethanol and was enhanced by the thioredoxin-reductase inhibitor auranofin. We predicted that Cys-119 or Cys-162 of p38α, close to the known MKK3 docking domain, were relevant for these redox characteristics. The C119S mutation decreased whereas the C162S mutation increased the dimer formation, suggesting that these two Cys residues act as vicinal thiols, consistent with C119S/C162S being incapable of sensing H2O2 Similarly, disulfide heterodimer formation was abolished in H9C2 cells expressing both MKK3 and p38α C119S/C162S and subjected to simulated ischemia and reperfusion. However, the p38α C119S/C162S mutants did not exhibit appreciable alteration in activating dual phosphorylation. In contrast, the anti-inflammatory agent 10-nitro-oleic acid (NO2-OA), a component of the Mediterranean diet, reduced p38α activation and covalently modified Cys-119/Cys-162, probably obstructing MKK3 access. Moreover, NO2-OA reduced the dephosphorylation of p38α by hematopoietic tyrosine phosphatase (HePTP). Furthermore, steric obstruction of Cys-119/Cys-162 by NO2-OA pretreatment in Langendorff-perfused murine hearts prevented the p38-MKK3 disulfide dimer formation and attenuated H2O2-induced contractile dysfunction. Our findings suggest that cysteine residues within p38α act as redox sensors that can dynamically regulate the association between p38 and MKK3.",
keywords = "Amino Acid Substitution, Animals, Cell Line, Cells, Cultured, Cysteine/chemistry, Cystine/chemistry, Enzyme Activation, Heart Ventricles/cytology, Humans, In Vitro Techniques, MAP Kinase Kinase 3/chemistry, Male, Mice, Inbred C57BL, Mitogen-Activated Protein Kinase 14/chemistry, Models, Molecular, Myocytes, Cardiac/cytology, Oxidation-Reduction, Oxidative Stress, Protein Conformation, Protein Multimerization, Rats, Wistar, Recombinant Fusion Proteins/chemistry",
author = "Rekha Bassi and Burgoyne, {Joseph R} and DeNicola, {Gian F} and Olena Rudyk and Vittorio DeSantis and Charles, {Rebecca L} and Philip Eaton and Marber, {Michael S}",
note = "{\textcopyright} 2017 by The American Society for Biochemistry and Molecular Biology, Inc.",
year = "2017",
month = sep,
day = "29",
doi = "10.1074/jbc.M117.785410",
language = "English",
volume = "292",
pages = "16161--16173",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "39",

}

RIS (suitable for import to EndNote) Download

TY - JOUR

T1 - Redox-dependent dimerization of p38α mitogen-activated protein kinase with mitogen-activated protein kinase kinase 3

AU - Bassi, Rekha

AU - Burgoyne, Joseph R

AU - DeNicola, Gian F

AU - Rudyk, Olena

AU - DeSantis, Vittorio

AU - Charles, Rebecca L

AU - Eaton, Philip

AU - Marber, Michael S

N1 - © 2017 by The American Society for Biochemistry and Molecular Biology, Inc.

PY - 2017/9/29

Y1 - 2017/9/29

N2 - The kinase p38α MAPK (p38α) plays a pivotal role in many biological processes. p38α is activated by canonical upstream kinases that phosphorylate the activation region. The purpose of our study was to determine whether such activation may depend on redox-sensing cysteines within p38α. p38α was activated and formed a disulfide-bound heterodimer with MAP2K3 (MKK3) in rat cardiomyocytes and isolated hearts exposed to H2O2 This disulfide heterodimer was sensitive to reduction by mercaptoethanol and was enhanced by the thioredoxin-reductase inhibitor auranofin. We predicted that Cys-119 or Cys-162 of p38α, close to the known MKK3 docking domain, were relevant for these redox characteristics. The C119S mutation decreased whereas the C162S mutation increased the dimer formation, suggesting that these two Cys residues act as vicinal thiols, consistent with C119S/C162S being incapable of sensing H2O2 Similarly, disulfide heterodimer formation was abolished in H9C2 cells expressing both MKK3 and p38α C119S/C162S and subjected to simulated ischemia and reperfusion. However, the p38α C119S/C162S mutants did not exhibit appreciable alteration in activating dual phosphorylation. In contrast, the anti-inflammatory agent 10-nitro-oleic acid (NO2-OA), a component of the Mediterranean diet, reduced p38α activation and covalently modified Cys-119/Cys-162, probably obstructing MKK3 access. Moreover, NO2-OA reduced the dephosphorylation of p38α by hematopoietic tyrosine phosphatase (HePTP). Furthermore, steric obstruction of Cys-119/Cys-162 by NO2-OA pretreatment in Langendorff-perfused murine hearts prevented the p38-MKK3 disulfide dimer formation and attenuated H2O2-induced contractile dysfunction. Our findings suggest that cysteine residues within p38α act as redox sensors that can dynamically regulate the association between p38 and MKK3.

AB - The kinase p38α MAPK (p38α) plays a pivotal role in many biological processes. p38α is activated by canonical upstream kinases that phosphorylate the activation region. The purpose of our study was to determine whether such activation may depend on redox-sensing cysteines within p38α. p38α was activated and formed a disulfide-bound heterodimer with MAP2K3 (MKK3) in rat cardiomyocytes and isolated hearts exposed to H2O2 This disulfide heterodimer was sensitive to reduction by mercaptoethanol and was enhanced by the thioredoxin-reductase inhibitor auranofin. We predicted that Cys-119 or Cys-162 of p38α, close to the known MKK3 docking domain, were relevant for these redox characteristics. The C119S mutation decreased whereas the C162S mutation increased the dimer formation, suggesting that these two Cys residues act as vicinal thiols, consistent with C119S/C162S being incapable of sensing H2O2 Similarly, disulfide heterodimer formation was abolished in H9C2 cells expressing both MKK3 and p38α C119S/C162S and subjected to simulated ischemia and reperfusion. However, the p38α C119S/C162S mutants did not exhibit appreciable alteration in activating dual phosphorylation. In contrast, the anti-inflammatory agent 10-nitro-oleic acid (NO2-OA), a component of the Mediterranean diet, reduced p38α activation and covalently modified Cys-119/Cys-162, probably obstructing MKK3 access. Moreover, NO2-OA reduced the dephosphorylation of p38α by hematopoietic tyrosine phosphatase (HePTP). Furthermore, steric obstruction of Cys-119/Cys-162 by NO2-OA pretreatment in Langendorff-perfused murine hearts prevented the p38-MKK3 disulfide dimer formation and attenuated H2O2-induced contractile dysfunction. Our findings suggest that cysteine residues within p38α act as redox sensors that can dynamically regulate the association between p38 and MKK3.

KW - Amino Acid Substitution

KW - Animals

KW - Cell Line

KW - Cells, Cultured

KW - Cysteine/chemistry

KW - Cystine/chemistry

KW - Enzyme Activation

KW - Heart Ventricles/cytology

KW - Humans

KW - In Vitro Techniques

KW - MAP Kinase Kinase 3/chemistry

KW - Male

KW - Mice, Inbred C57BL

KW - Mitogen-Activated Protein Kinase 14/chemistry

KW - Models, Molecular

KW - Myocytes, Cardiac/cytology

KW - Oxidation-Reduction

KW - Oxidative Stress

KW - Protein Conformation

KW - Protein Multimerization

KW - Rats, Wistar

KW - Recombinant Fusion Proteins/chemistry

UR - http://www.scopus.com/inward/record.url?scp=85030032567&partnerID=8YFLogxK

U2 - 10.1074/jbc.M117.785410

DO - 10.1074/jbc.M117.785410

M3 - Article

C2 - 28739872

AN - SCOPUS:85030032567

VL - 292

SP - 16161

EP - 16173

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 39

ER -

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