期刊
EXPERIMENTAL GERONTOLOGY
卷 44, 期 6-7, 页码 375-382出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.exger.2009.02.009
关键词
Lamin A/C; Oxidative stress; SH-SY5Y cell; 6-Hydroxy dopamine; Blue-native electrophoresis; Proteomics
资金
- Japan Society for the Promotion of Science
There is accumulating evidence that oxidative stress plays an important role in aging. Our previous phosphoproteomic study of the human neuroblastoma cell line SH-SY5Y revealed changes in the phosphorylation of several proteins such as lamin A/C during 6-hydroxydopamine-induced oxidative stress. The present study employed native proteomic analysis to clarify protein-protein interaction under physiological conditions. We examined oxidative stress-related changes in SH-SY5Y cellular proteins using blue-native polyacrylamide gel electrophoresis (BN-PAGE), a powerful tool for the separation of protein complexes. BN-PAGE gel images showed successful separation of several complexes. Components of these complexes, separated by 2-D BN-PAGE in combination with SDS-PAGE, were identified by peptide mass fingerprinting employing MALDI-TOF MS and an MS/MS ion search on LC-MS/MS. TCP-1 complex, ATP synthase, and the complex of heat shock protein 90 with its client proteins such as pyruvate kinase were detected. Two dimensional BN-PAGE and Western blot analysis revealed an increase of lamin A/C associated with heat shock protein 90 in response to 6-OHDA-induced oxidative stress. (C) 2009 Elsevier Inc. All rights reserved.
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