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Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment
Recent advancements in isolation techniques for cytochrome c (Cytc) have allowed us to discover post-translational modifications of this protein. We previously identified two distinct tyrosine phosphorylated residues on Cytc in mammalian liver and heart that alter its electron transfer kinetics and...
Autores principales: | , , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2013
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3818486/ https://www.ncbi.nlm.nih.gov/pubmed/24223835 http://dx.doi.org/10.1371/journal.pone.0078627 |
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author | Sanderson, Thomas H. Mahapatra, Gargi Pecina, Petr Ji, Qinqin Yu, Kebing Sinkler, Christopher Varughese, Ashwathy Kumar, Rita Bukowski, Melissa J. Tousignant, Renee N. Salomon, Arthur R. Lee, Icksoo Hüttemann, Maik |
author_facet | Sanderson, Thomas H. Mahapatra, Gargi Pecina, Petr Ji, Qinqin Yu, Kebing Sinkler, Christopher Varughese, Ashwathy Kumar, Rita Bukowski, Melissa J. Tousignant, Renee N. Salomon, Arthur R. Lee, Icksoo Hüttemann, Maik |
author_sort | Sanderson, Thomas H. |
collection | PubMed |
description | Recent advancements in isolation techniques for cytochrome c (Cytc) have allowed us to discover post-translational modifications of this protein. We previously identified two distinct tyrosine phosphorylated residues on Cytc in mammalian liver and heart that alter its electron transfer kinetics and the ability to induce apoptosis. Here we investigated the phosphorylation status of Cytc in ischemic brain and sought to determine if insulin-induced neuroprotection and inhibition of Cytc release was associated with phosphorylation of Cytc. Using an animal model of global brain ischemia, we found a ∼50% decrease in neuronal death in the CA1 hippocampal region with post-ischemic insulin administration. This insulin-mediated increase in neuronal survival was associated with inhibition of Cytc release at 24 hours of reperfusion. To investigate possible changes in the phosphorylation state of Cytc we first isolated the protein from ischemic pig brain and brain that was treated with insulin. Ischemic brains demonstrated no detectable tyrosine phosphorylation. In contrast Cytc isolated from brains treated with insulin showed robust phosphorylation of Cytc, and the phosphorylation site was unambiguously identified as Tyr97 by immobilized metal affinity chromatography/nano-liquid chromatography/electrospray ionization mass spectrometry. We next confirmed these results in rats by in vivo application of insulin in the absence or presence of global brain ischemia and determined that Cytc Tyr97-phosphorylation is strongly induced under both conditions but cannot be detected in untreated controls. These data suggest a mechanism whereby Cytc is targeted for phosphorylation by insulin signaling, which may prevent its release from the mitochondria and the induction of apoptosis. |
format | Online Article Text |
id | pubmed-3818486 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2013 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-38184862013-11-09 Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment Sanderson, Thomas H. Mahapatra, Gargi Pecina, Petr Ji, Qinqin Yu, Kebing Sinkler, Christopher Varughese, Ashwathy Kumar, Rita Bukowski, Melissa J. Tousignant, Renee N. Salomon, Arthur R. Lee, Icksoo Hüttemann, Maik PLoS One Research Article Recent advancements in isolation techniques for cytochrome c (Cytc) have allowed us to discover post-translational modifications of this protein. We previously identified two distinct tyrosine phosphorylated residues on Cytc in mammalian liver and heart that alter its electron transfer kinetics and the ability to induce apoptosis. Here we investigated the phosphorylation status of Cytc in ischemic brain and sought to determine if insulin-induced neuroprotection and inhibition of Cytc release was associated with phosphorylation of Cytc. Using an animal model of global brain ischemia, we found a ∼50% decrease in neuronal death in the CA1 hippocampal region with post-ischemic insulin administration. This insulin-mediated increase in neuronal survival was associated with inhibition of Cytc release at 24 hours of reperfusion. To investigate possible changes in the phosphorylation state of Cytc we first isolated the protein from ischemic pig brain and brain that was treated with insulin. Ischemic brains demonstrated no detectable tyrosine phosphorylation. In contrast Cytc isolated from brains treated with insulin showed robust phosphorylation of Cytc, and the phosphorylation site was unambiguously identified as Tyr97 by immobilized metal affinity chromatography/nano-liquid chromatography/electrospray ionization mass spectrometry. We next confirmed these results in rats by in vivo application of insulin in the absence or presence of global brain ischemia and determined that Cytc Tyr97-phosphorylation is strongly induced under both conditions but cannot be detected in untreated controls. These data suggest a mechanism whereby Cytc is targeted for phosphorylation by insulin signaling, which may prevent its release from the mitochondria and the induction of apoptosis. Public Library of Science 2013-11-05 /pmc/articles/PMC3818486/ /pubmed/24223835 http://dx.doi.org/10.1371/journal.pone.0078627 Text en © 2013 Sanderson et al http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Sanderson, Thomas H. Mahapatra, Gargi Pecina, Petr Ji, Qinqin Yu, Kebing Sinkler, Christopher Varughese, Ashwathy Kumar, Rita Bukowski, Melissa J. Tousignant, Renee N. Salomon, Arthur R. Lee, Icksoo Hüttemann, Maik Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title | Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title_full | Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title_fullStr | Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title_full_unstemmed | Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title_short | Cytochrome c Is Tyrosine 97 Phosphorylated by Neuroprotective Insulin Treatment |
title_sort | cytochrome c is tyrosine 97 phosphorylated by neuroprotective insulin treatment |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3818486/ https://www.ncbi.nlm.nih.gov/pubmed/24223835 http://dx.doi.org/10.1371/journal.pone.0078627 |
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