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Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury

Ischemia/reperfusion injury (IRI) occurs inevitably in liver transplantations and frequently during major resections, and can lead to liver dysfunction as well as systemic disorders. High-mobility group box 1 (HMGB1) plays a pathogenic role in hepatic IRI. In the normal liver, HMGB1 is located in th...

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Autores principales: Zhang, Tianjiao, Wei, Weiwei, Dirsch, Olaf, Krüger, Thomas, Kan, Chunyi, Xie, Chichi, Kniemeyer, Olaf, Fang, Haoshu, Settmacher, Utz, Dahmen, Uta
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297800/
https://www.ncbi.nlm.nih.gov/pubmed/28275217
http://dx.doi.org/10.3390/ijms18010167
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author Zhang, Tianjiao
Wei, Weiwei
Dirsch, Olaf
Krüger, Thomas
Kan, Chunyi
Xie, Chichi
Kniemeyer, Olaf
Fang, Haoshu
Settmacher, Utz
Dahmen, Uta
author_facet Zhang, Tianjiao
Wei, Weiwei
Dirsch, Olaf
Krüger, Thomas
Kan, Chunyi
Xie, Chichi
Kniemeyer, Olaf
Fang, Haoshu
Settmacher, Utz
Dahmen, Uta
author_sort Zhang, Tianjiao
collection PubMed
description Ischemia/reperfusion injury (IRI) occurs inevitably in liver transplantations and frequently during major resections, and can lead to liver dysfunction as well as systemic disorders. High-mobility group box 1 (HMGB1) plays a pathogenic role in hepatic IRI. In the normal liver, HMGB1 is located in the nucleus of hepatocytes; after ischemia reperfusion, it translocates to the cytoplasm and it is further released to the extracellular space. Unlike the well-explored functions of nuclear and extracellular HMGB1, the role of cytoplasmic HMGB1 in hepatic IRI remains elusive. We hypothesized that cytoplasmic HMGB1 interacts with binding proteins involved in the hepatocellular response to IRI. In this study, binding proteins of cytoplasmic HMGB1 during hepatic IRI were identified. Liver tissues from rats with warm ischemia reperfusion (WI/R) injury and from normal rats were subjected to cytoplasmic protein extraction. Co-immunoprecipitation using these protein extracts was performed to enrich HMGB1-protein complexes. To separate and identify the immunoprecipitated proteins in eluates, 2-dimensional electrophoresis and subsequent mass spectrometry detection were performed. Two of the identified proteins were verified using Western blotting: betaine–homocysteine S-methyltransferase 1 (BHMT) and cystathionine γ-lyase (CTH). Therefore, our results revealed the binding of HMGB1 to BHMT and CTH in cytoplasm during hepatic WI/R. This finding may help to better understand the cellular response to IRI in the liver and to identify novel molecular targets for reducing ischemic injury.
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spelling pubmed-52978002017-02-10 Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury Zhang, Tianjiao Wei, Weiwei Dirsch, Olaf Krüger, Thomas Kan, Chunyi Xie, Chichi Kniemeyer, Olaf Fang, Haoshu Settmacher, Utz Dahmen, Uta Int J Mol Sci Article Ischemia/reperfusion injury (IRI) occurs inevitably in liver transplantations and frequently during major resections, and can lead to liver dysfunction as well as systemic disorders. High-mobility group box 1 (HMGB1) plays a pathogenic role in hepatic IRI. In the normal liver, HMGB1 is located in the nucleus of hepatocytes; after ischemia reperfusion, it translocates to the cytoplasm and it is further released to the extracellular space. Unlike the well-explored functions of nuclear and extracellular HMGB1, the role of cytoplasmic HMGB1 in hepatic IRI remains elusive. We hypothesized that cytoplasmic HMGB1 interacts with binding proteins involved in the hepatocellular response to IRI. In this study, binding proteins of cytoplasmic HMGB1 during hepatic IRI were identified. Liver tissues from rats with warm ischemia reperfusion (WI/R) injury and from normal rats were subjected to cytoplasmic protein extraction. Co-immunoprecipitation using these protein extracts was performed to enrich HMGB1-protein complexes. To separate and identify the immunoprecipitated proteins in eluates, 2-dimensional electrophoresis and subsequent mass spectrometry detection were performed. Two of the identified proteins were verified using Western blotting: betaine–homocysteine S-methyltransferase 1 (BHMT) and cystathionine γ-lyase (CTH). Therefore, our results revealed the binding of HMGB1 to BHMT and CTH in cytoplasm during hepatic WI/R. This finding may help to better understand the cellular response to IRI in the liver and to identify novel molecular targets for reducing ischemic injury. MDPI 2017-01-16 /pmc/articles/PMC5297800/ /pubmed/28275217 http://dx.doi.org/10.3390/ijms18010167 Text en © 2017 by the authors; licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC-BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Zhang, Tianjiao
Wei, Weiwei
Dirsch, Olaf
Krüger, Thomas
Kan, Chunyi
Xie, Chichi
Kniemeyer, Olaf
Fang, Haoshu
Settmacher, Utz
Dahmen, Uta
Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title_full Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title_fullStr Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title_full_unstemmed Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title_short Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury
title_sort identification of proteins interacting with cytoplasmic high-mobility group box 1 during the hepatocellular response to ischemia reperfusion injury
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5297800/
https://www.ncbi.nlm.nih.gov/pubmed/28275217
http://dx.doi.org/10.3390/ijms18010167
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