Analysis of oxygen/glucose-deprivation-induced changes in SUMO3 conjugation using SILAC-based quantitative proteomics.
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.
Type
Journal articleSubject
Cell Line, TumorAnimals
Mice
Neuroblastoma
Oxygen
Glucose
Proteins
Small Ubiquitin-Related Modifier Proteins
Isotope Labeling
Proteomics
Cell Hypoxia
Ubiquitination
Stress, Physiological
Protein Interaction Maps
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https://hdl.handle.net/10161/23288Published Version (Please cite this version)
10.1021/pr200834fPublication Info
Yang, W; Thompson, JW; Wang, Z; Wang, L; Sheng, H; Foster, MW; ... Paschen, W (2012). Analysis of oxygen/glucose-deprivation-induced changes in SUMO3 conjugation using
SILAC-based quantitative proteomics. Journal of proteome research, 11(2). pp. 1108-1117. 10.1021/pr200834f. Retrieved from https://hdl.handle.net/10161/23288.This is constructed from limited available data and may be imprecise. To cite this
article, please review & use the official citation provided by the journal.
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Show full item recordScholars@Duke
Matthew Wolf Foster
Associate Professor in Medicine
Martin Arthur Moseley III
Adjunct Professor in the Department of Cell Biology
Wulf Paschen
Professor in Anesthesiology
My research interests are understanding the mechanisms underlying induction of cell
death induced by a severe form of cellular stress. I am particularly interested in
the role of the endoplasmic reticulum in the pathological process induced by transient
cerebral ischemia and culminating in neuronal cell death. This pathological process
is associated with an irreversible suppression of protein synthese that limits the
ability of cells to withstand ischemia-induced impairment of endoplasmic r
Huaxin Sheng
Associate Professor in Anesthesiology
We have successfully developed various rodent models of brain and spinal cord injuries
in our lab, such as focal cerebral ischemia, global cerebral ischemia, head trauma,
subarachnoid hemorrhage, intracerebral hemorrhage, spinal cord ischemia and compression
injury. We also established cardiac arrest and hemorrhagic shock models for studying
multiple organ dysfunction. Our current studies focus on two projects. One is to
examine the efficacy of catalytic antioxidant in treating cerebral is
J. Will Thompson
Adjunct Assistant Professor in the Department of Pharmacology & Cancer Biology
Dr. Thompson's research focuses on the development and deployment of proteomics and
metabolomics mass spectrometry techniques for the analysis of biological systems.
He served as the Assistant Director of the Proteomics and Metabolomics Shared Resource
in the Duke School of Medicine from 2007-2021. He currently maintains collaborations
in metabolomics and proteomics research at Duke, and develops new tools for chemical
analysis as a Princi
Wei Yang
Associate Professor in Anesthesiology
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