title: Phosphoproteomic analysis of neuronal cell death by glutamate-induced oxidative stress.
abstract: 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.
authors.affiliation: Translational Research Center, KRIBB, Daejeon, South Korea.
authors.forename: Tae Hyuk
authors.initials: TH
authors.lastname: Kang
authors.forename: Kwang-Hee
authors.initials: KH
authors.lastname: Bae
authors.forename: Min-Jung
authors.initials: MJ
authors.lastname: Yu
authors.forename: Won-Kon
authors.initials: WK
authors.lastname: Kim
authors.forename: Hyang-Ran
authors.initials: HR
authors.lastname: Hwang
authors.forename: Hyeyun
authors.initials: H
authors.lastname: Jung
authors.forename: Phil Young
authors.initials: PY
authors.lastname: Lee
authors.forename: Sunghyun
authors.initials: S
authors.lastname: Kang
authors.forename: Tae-Sung
authors.initials: TS
authors.lastname: Yoon
authors.forename: Sung Goo
authors.initials: SG
authors.lastname: Park
authors.forename: Seong Eon
authors.initials: SE
authors.lastname: Ryu
authors.forename: Sang Chul
authors.initials: SC
authors.lastname: Lee
mesh_terms: 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
chemical_list: D010750:Phosphoproteins; D020543:Proteome; D018698:Glutamic Acid; D004533:Egtazic Acid; D004492:Edetic Acid