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Age‐dependent variations of cell response to oxidative stress: Proteomic approach to protein expression and phosphorylation
Author(s) -
Miura Yuri,
Kano Mayumi,
Abe Kazuhiko,
Urano Shiro,
Suzuki Shozo,
Toda Tosifusa
Publication year - 2005
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.200500172
Subject(s) - oxidative stress , protein expression , phosphorylation , oxidative phosphorylation , proteomics , microbiology and biotechnology , chemistry , cell , fight or flight response , expression (computer science) , biology , biochemistry , biophysics , computer science , gene , programming language
We investigated the protein profiles of variously aged rat astrocytes in response to oxidative stress. After H 2 O 2 ‐exposure of cells at 100 µ M for 30 min, the relative intensity of ten protein spots changed on two‐dimensional (2‐D) gels compared with control gels after silver staining. Matrix‐assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) analysis after in‐gel digestion revealed that six of these spots corresponded to three kinds of proteins, each of which was composed of a protein and its modified form with a different isoelectric point (p I ). These three proteins were identified as peroxiredoxins (PRDXs) II and III, and calpactin I light chain (p11). H 2 O 2 ‐exposure increased the intensity of the spot with lower p I and simultaneously decreased that of the spot with higher p I for both PRDXs II and III. In addition, the expression of annexin VII, S ‐adenosyl‐ L ‐homocysteine hydrolase, elongation factor II fragment (EF‐II), and adenosine deaminase was increased by H 2 O 2 ‐exposure in astrocytes from variously aged rats. Using the Pro‐Q® Diamond staining, heat shock protein 60 kDa (Hsp 60) and α‐tubulin were observed to be phosphorylated upon H 2 O 2 ‐exposure. While phosphorylation of α‐tubulin was correlated positively with age, the changes in abundance of ten protein spots as described above were independent of age. These results suggest that aging does not suppress the responses aimed at limiting injury and promoting repair brought about by severe oxidative stress, and might affect cell dynamics including the formation of microtubules.