Phosphoproteomic analysis of neuronal cell death by glutamate-induced oxidative stress.
Oxidative stress is one of the major causes of neuronal cell death in disorders such as perinatal hypoxia and ischemia. Protein phosphorylation is the most significant PTM of proteins and plays an important role in stress-induced signal transduction. Thus, the analysis of alternative protein phosphorylation states which occur during oxidative stress-induced cell death could provide valuable information regarding cell death. In this study, a reference phosphoproteome map of the mouse hippocampal cell line HT22 was constructed based on 125 spots that were identified by MALDI-TOF or LC-ESI-Q-TOF-MS analysis. In addition, proteins of HT22 cells at various stages of oxidative stress-induced cell death were separated by 2-DE and alterations in phosphoproteins were detected by Pro-Q Diamond staining. A total of 17 spots showing significant quantitative changes and seven newly appearing spots were identified after glutamate treatment. Splicing factor 2, peroxiredoxin 2, S100 calcium binding protein A11, and purine nucleoside phosphorylase were identified as up- or down-regulated proteins. CDC25A, caspase-8, and cyp51 protein appeared during oxidative stress-induced cell death. The data in this study from phosphoproteomic analysis provide a valuable resource for the understanding of HT22 cell death mechanisms mediated by oxidative stress.
Kang
Tae Hyuk
TH
Translational Research Center, KRIBB, Daejeon, South Korea.
Bae
Kwang-Hee
KH
Yu
Min-Jung
MJ
Kim
Won-Kon
WK
Hwang
Hyang-Ran
HR
Jung
Hyeyun
H
Lee
Phil Young
PY
Kang
Sunghyun
S
Yoon
Tae-Sung
TS
Park
Sung Goo
SG
Ryu
Seong Eon
SE
Lee
Sang Chul
SC
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D016923:Cell Death / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D004492:Edetic Acid / Q000494:pharmacology; D004533:Egtazic Acid / Q000494:pharmacology; D018698:Glutamic Acid / Q000494:pharmacology*; D006624:Hippocampus / Q000166:cytology; D013058:Mass Spectrometry; D051379:Mice; D009474:Neurons / Q000187:drug effects* / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D010750:Phosphoproteins / Q000032:analysis / Q000378:metabolism; D010766:Phosphorylation; D020543:Proteome / Q000032:analysis*; D040901:Proteomics / Q000379:methods; D013997:Time Factors
D010750:Phosphoproteins; D020543:Proteome; D018698:Glutamic Acid; D004533:Egtazic Acid; D004492:Edetic Acid