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 | |
dc.contributor.author | Wang, Z | |
dc.contributor.author | Wang, L | |
dc.contributor.author | Sheng, H | |
dc.contributor.author | Foster, MW | |
dc.contributor.author | Moseley, MA | |
dc.contributor.author | Paschen, W | |
dc.date.accessioned | 2021-06-01T14:14:36Z | |
dc.date.available | 2021-06-01T14:14:36Z | |
dc.date.issued | 2012-02 | |
dc.date.updated | 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 | |
dc.identifier.issn | 1535-3907 | |
dc.identifier.uri | ||
dc.language | eng | |
dc.publisher | American Chemical Society (ACS) | |
dc.relation.ispartof | Journal of proteome research | |
dc.relation.isversionof | 10.1021/pr200834f | |
dc.subject | Cell Line, Tumor | |
dc.subject | Animals | |
dc.subject | Mice | |
dc.subject | Neuroblastoma | |
dc.subject | Oxygen | |
dc.subject | Glucose | |
dc.subject | Proteins | |
dc.subject | Small Ubiquitin-Related Modifier Proteins | |
dc.subject | Isotope Labeling | |
dc.subject | Proteomics | |
dc.subject | Cell Hypoxia | |
dc.subject | Ubiquitination | |
dc.subject | Stress, Physiological | |
dc.subject | 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 | |
pubs.organisational-group | Medicine, Pulmonary, Allergy, and Critical Care Medicine | |
pubs.organisational-group | Duke | |
pubs.organisational-group | Medicine | |
pubs.organisational-group | Clinical Science Departments | |
pubs.organisational-group | Medicine, Cardiology | |
pubs.organisational-group | Faculty | |
pubs.organisational-group | Anesthesiology | |
pubs.organisational-group | Pharmacology & Cancer Biology | |
pubs.organisational-group | Basic Science Departments | |
pubs.publication-status | Published | |
pubs.volume | 11 |