Characterization of the ubiquitin-modified proteome regulated by transient forebrain ischemia.
Abstract
Ubiquitylation is a posttranslational protein modification that modulates various
cellular processes of key significance, including protein degradation and DNA damage
repair. In animals subjected to transient cerebral ischemia, ubiquitin-conjugated
proteins accumulate in Triton-insoluble aggregates. Although this process is widely
considered to modulate the fate of postischemic neurons, few attempts have been made
to characterize the ubiquitin-modified proteome in these aggregates. We performed
proteomics analyses to identify ubiquitylated proteins in postischemic aggregates.
Mice were subjected to 10 minutes of forebrain ischemia and 4 hours of reperfusion.
The hippocampi were dissected, aggregates were isolated, and trypsin-digested after
spiking with GG-BSA as internal standard. K-ɛ-GG-containing peptides were immunoprecipitated
and analyzed by label-free quantitative liquid chromatography tandem mass spectrometry
(LC-MS/MS) analysis. We identified 1,664 peptides to 520 proteins containing at least
one K-ɛ-GG. Sixty-six proteins were highly ubiquitylated, with 10 or more K-ɛ-GG peptides.
Based on selection criteria of greater than fivefold increase and P<0.001, 763 peptides
to 272 proteins were highly enriched in postischemic aggregates. These included proteins
involved in important neuronal functions and signaling pathways that are impaired
after ischemia. Results of this study could serve as an important platform to uncover
the mechanisms linking insoluble ubiquitin aggregates to the functions of postischemic
neurons.
Type
Journal articleSubject
ProsencephalonAnimals
Mice, Inbred C57BL
Mice
Ischemic Attack, Transient
Peptide Fragments
Proteome
Ubiquitin
Microscopy, Confocal
Blotting, Western
Chromatography, Liquid
Proteomics
Male
Tandem Mass Spectrometry
Ubiquitination
Ubiquitinated Proteins
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https://hdl.handle.net/10161/23272Published Version (Please cite this version)
10.1038/jcbfm.2013.210Publication Info
Iwabuchi, Masahiro; Sheng, Huaxin; Thompson, J Will; Wang, Liangli; Dubois, Laura
G; Gooden, David; ... Yang, Wei (2014). Characterization of the ubiquitin-modified proteome regulated by transient forebrain
ischemia. Journal of cerebral blood flow and metabolism : official journal of the International
Society of Cerebral Blood Flow and Metabolism, 34(3). pp. 425-432. 10.1038/jcbfm.2013.210. Retrieved from https://hdl.handle.net/10161/23272.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
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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