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Functional impact of PTP1B-mediated Src regulation on oxidative phosphorylation in rat brain mitochondria

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Abstract

Given the presence of Src and PTP1B within rat brain mitochondria, we have investigated whether PTP1B regulates Src activity in mitochondria as in the cytosol. Results showed that Src was stimulated by in vitro addition of ATP to mitochondria, and this stimulation was reversed by a membrane-permeable allosteric inhibitor of PTP1B and by a potent selective Src inhibitor. They also indicated a direct action of PTP1B on phosphorylated tyrosine 527 residue of Src, thus implicating a role for PTP1B in the modulation of Src activity in mitochondria. Putative Src and PTP1B substrates were identified by liquid chromatography tandem mass spectrometry and two-dimensional blue native/SDS-PAGE. Both inhibitors inhibited ADP-stimulated respirations concurrently with Src activation and complex IV activation by ATP, while having no effect or increasing the activity of the other complexes. Our analysis emphasizes the regulatory function of Src and its modulation by PTP1B on oxidative phosphorylation in mitochondria.

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Fig. 1
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Fig. 5a, b
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Abbreviations

OxPhos:

Oxidative phosphorylation

Y:

Tyrosine

Csk:

C terminal Src kinase

PP2:

4-Amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine

PTP1Bi:

3-(3,5-Dibromo-4-hydroxy-benzoyl)-2-ethyl-benzofuran-6-sulfonic acid-(4-(thiazol-2-ylsulfamyl)-phenyl)-amide inhibitor

2D BN/SDS-PAGE:

Two-dimensional blue native/sodium dodecyl sulfate polyacrylamide gel electrophoresis

Nano LC-MS/MS:

Nano liquid chromatography tandem mass spectrometry

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Acknowledgments

We thank Nadia Barrier for her technical support. Etienne Hébert Chatelain was financially supported by FQRNT (Fonds Québécois de la Recherche sur la Nature et les Technologies) and NSERC (National Sciences and Engineering Resaerch Council of Canada) scholarships. This work was supported in part by grants from INSERM, the University Victor Segalen-Bordeaux 2, and the Fondation de France (N°Engt 2008002720).

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Authors and Affiliations

  1. Physiopathologie Mitochondriale, INSERM-U688, Université Victor Ségalen-Bordeaux 2, 146 rue Léo Saignat, 33076, Bordeaux-Cedex, France

    Etienne Hébert Chatelain, Thierry Letellier & Jeanne Dachary-Prigent

  2. Pôle Protéomique, Centre de Génomique Fonctionnelle, Université de Bordeaux, 146 rue Léo Saignat, 33076, Bordeaux, France

    Jean-William Dupuy

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  1. Etienne Hébert Chatelain
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Correspondence to Etienne Hébert Chatelain.

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Hébert Chatelain, E., Dupuy, JW., Letellier, T. et al. Functional impact of PTP1B-mediated Src regulation on oxidative phosphorylation in rat brain mitochondria. Cell. Mol. Life Sci. 68, 2603–2613 (2011). https://doi.org/10.1007/s00018-010-0573-6

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