Proteomic identification of carbonylated proteins in the monkey hippocampus after ischemia-reperfusion.
Reactive oxygen species (ROS) are known to participate in neurodegeneration after ischemia-reperfusion. With the aid of ROS, the calpain-induced lysosomal rupture provokes ischemic neuronal death in the cornu Ammonis (CA) 1 of the hippocampus; however, the target proteins of ROS still remain unknown. Here a proteomic analysis was done to identify and characterize ROS-induced carbonyl modification of proteins in the CA1 of the macaque monkey after transient whole-brain ischemia followed by reperfusion. We found that carbonyl modification of heat shock 70-kDa protein 1 (Hsp70-1), a major stress-inducible member of the Hsp70 family, was extensively increased before the neuronal death in the CA1 sector, and the carbonylation site was identified to be Arg469 of Hsp70-1. The CA1 neuronal death conceivably occurs by calpain-mediated cleavage of carbonylated Hsp70 that becomes prone to proteolysis with the resultant lysosomal rupture. In addition, the carbonyl levels of dihydropyrimidinase-like 2 isoform 2, glial fibrillary acidic protein, and beta-actin were remarkably increased in the postischemic CA1. Therefore, ischemia-reperfusion-induced oxidative damage to these proteins in the CA1 may lead to loss of the neuroprotective function, which contributes to neuronal death.
Oikawa
Shinji
S
Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Mie 514-8507, Japan. s-oikawa@doc.medic.mie-u.ac.jp
Yamada
Tomoko
T
Minohata
Toshikazu
T
Kobayashi
Hatasu
H
Furukawa
Ayako
A
Tada-Oikawa
Saeko
S
Hiraku
Yusuke
Y
Murata
Mariko
M
Kikuchi
Mitsuru
M
Yamashima
Tetsumori
T
D000199:Actins / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D002154:Calpain / Q000378:metabolism; D016923:Cell Death; D005904:Glial Fibrillary Acidic Protein / Q000235:genetics / Q000378:metabolism*; D050884:HSP72 Heat-Shock Proteins / Q000378:metabolism*; D006624:Hippocampus; D036341:Intercellular Signaling Peptides and Proteins / Q000235:genetics / Q000378:metabolism*; D008251:Macaca; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism* / Q000473:pathology; D050050:Protein Carbonylation; D020033:Protein Isoforms; D040901:Proteomics; D017382:Reactive Oxygen Species; D015427:Reperfusion Injury
D000199:Actins; D005904:Glial Fibrillary Acidic Protein; D050884:HSP72 Heat-Shock Proteins; D036341:Intercellular Signaling Peptides and Proteins; D009419:Nerve Tissue Proteins; D020033:Protein Isoforms; D017382:Reactive Oxygen Species; C095114:collapsin response mediator protein-2; D002154:Calpain