Analysis of oxygen/glucose-deprivation-induced changes in SUMO3 conjugation using SILAC-based quantitative proteomics.

dc.contributor.author

Yang, W

dc.contributor.author

Thompson, JW

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Wang, Z

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Wang, L

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Sheng, H

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Foster, MW

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Moseley, MA

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Paschen, W

dc.date.accessioned

2021-06-01T14:14:36Z

dc.date.available

2021-06-01T14:14:36Z

dc.date.issued

2012-02

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2021-06-01T14:14:36Z

dc.description.abstract

Transient cerebral ischemia dramatically activates small ubiquitin-like modifier (SUMO2/3) conjugation. In cells exposed to 6 h of transient oxygen/glucose deprivation (OGD), a model of ischemia, SUMOylation increases profoundly between 0 and 30 min following re-oxygenation. To elucidate the effect of transient OGD on SUMO conjugation of target proteins, we exposed neuroblastoma B35 cells expressing HA-SUMO3 to transient OGD and used stable isotope labeling with amino acids in cell culture (SILAC) to quantify OGD-induced changes in levels of specific SUMOylated proteins. Lysates from control and OGD-treated cells were mixed equally, and HA-tagged proteins were immunoprecipitated and analyzed by 1D-SDS-PAGE-LC-MS/MS. We identified 188 putative SUMO3-conjugated proteins, including numerous transcription factors and coregulators, and PIAS2 and PIAS4 SUMO ligases, of which 22 were increased or decreased more than ±2-fold. In addition to SUMO3, the levels of protein-conjugated SUMO1 and SUMO2, as well as ubiquitin, were all increased. Importantly, protein ubiquitination induced by OGD was completely blocked by gene silencing of SUMO2/3. Collectively, these results suggest several mechanisms for OGD-modulated SUMOylation, point to a number of signaling pathways that may be targets of SUMO-based signaling and recovery from ischemic stress, and demonstrate a tightly controlled crosstalk between the SUMO and ubiquitin conjugation pathways.

dc.identifier.issn

1535-3893

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1535-3907

dc.identifier.uri

https://hdl.handle.net/10161/23288

dc.language

eng

dc.publisher

American Chemical Society (ACS)

dc.relation.ispartof

Journal of proteome research

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10.1021/pr200834f

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Cell Line, Tumor

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Animals

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Mice

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Neuroblastoma

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Oxygen

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Glucose

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Proteins

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Small Ubiquitin-Related Modifier Proteins

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Isotope Labeling

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Proteomics

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Cell Hypoxia

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Ubiquitination

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Stress, Physiological

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Protein Interaction Maps

dc.title

Analysis of oxygen/glucose-deprivation-induced changes in SUMO3 conjugation using SILAC-based quantitative proteomics.

dc.type

Journal article

duke.contributor.orcid

Yang, W|0000-0001-5719-4393

duke.contributor.orcid

Sheng, H|0000-0002-4325-2940

duke.contributor.orcid

Foster, MW|0000-0003-0212-2346

pubs.begin-page

1108

pubs.end-page

1117

pubs.issue

2

pubs.organisational-group

School of Medicine

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Medicine, Pulmonary, Allergy, and Critical Care Medicine

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Duke

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Medicine

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Clinical Science Departments

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Medicine, Cardiology

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Faculty

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Anesthesiology

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Pharmacology & Cancer Biology

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Basic Science Departments

pubs.publication-status

Published

pubs.volume

11

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