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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MDPI AG
2017-01-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | http://www.mdpi.com/1422-0067/18/1/167 |
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| author | Tianjiao Zhang Weiwei Wei Olaf Dirsch Thomas Krüger Chunyi Kan Chichi Xie Olaf Kniemeyer Haoshu Fang Utz Settmacher Uta Dahmen |
| author_facet | Tianjiao Zhang Weiwei Wei Olaf Dirsch Thomas Krüger Chunyi Kan Chichi Xie Olaf Kniemeyer Haoshu Fang Utz Settmacher Uta Dahmen |
| author_sort | Tianjiao Zhang |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-11T23:42:50Z |
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| institution | Directory Open Access Journal |
| issn | 1422-0067 |
| language | English |
| last_indexed | 2024-04-11T23:42:50Z |
| publishDate | 2017-01-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-f674d73c5899475db8d814aaa8fa65932022-12-22T03:56:45ZengMDPI AGInternational Journal of Molecular Sciences1422-00672017-01-0118116710.3390/ijms18010167ijms18010167Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion InjuryTianjiao Zhang0Weiwei Wei1Olaf Dirsch2Thomas Krüger3Chunyi Kan4Chichi Xie5Olaf Kniemeyer6Haoshu Fang7Utz Settmacher8Uta Dahmen9Experimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyExperimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyInstitute of Pathology, Klinikum Chemnitz gGmbH, 09116 Chemnitz, GermanyDepartment of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institute, 07745 Jena, GermanyExperimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyExperimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyDepartment of Molecular and Applied Microbiology, Leibniz Institute for Natural Product Research and Infection Biology, Hans Knöll Institute, 07745 Jena, GermanyDepartment of Pathophysiology, Anhui Medical University, Hefei 230032, ChinaExperimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyExperimental Transplantation Surgery, Department of General, Visceral and Vascular Surgery, Jena University Hospital, 07747 Jena, GermanyIschemia/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.http://www.mdpi.com/1422-0067/18/1/1672-dimensional electrophoresis (2DE)ischemic damage responsehigh-mobility group box 1 (HMGB1)ischemic injurymass spectrometry (MS) |
| spellingShingle | Tianjiao Zhang Weiwei Wei Olaf Dirsch Thomas Krüger Chunyi Kan Chichi Xie Olaf Kniemeyer Haoshu Fang Utz Settmacher Uta Dahmen Identification of Proteins Interacting with Cytoplasmic High-Mobility Group Box 1 during the Hepatocellular Response to Ischemia Reperfusion Injury International Journal of Molecular Sciences 2-dimensional electrophoresis (2DE) ischemic damage response high-mobility group box 1 (HMGB1) ischemic injury mass spectrometry (MS) |
| 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 | 2-dimensional electrophoresis (2DE) ischemic damage response high-mobility group box 1 (HMGB1) ischemic injury mass spectrometry (MS) |
| url | http://www.mdpi.com/1422-0067/18/1/167 |
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