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| Serine Protease Omi/HtrA2 Regulates Adaptor Protein p66Shc in Mitochondria |
| LIU Shu1,2, JIANG Chang-an 1,2 |
1. West China Development and Stem Cell Institute, West China Institute of Women and Children's Health, West China Second University Hospital, Chengdu 610041, China;
2. State Key Laboratory of Biotherapy, West China Hospital, Chengdu 610041, China |
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Abstract Objective: Both accumulation of p66Shc in mitochondria and loss of Omi/HtrA2 lead to mitochondrial dysfunction, which triggers apoptosis. To study the regulation of HtrA2 to the mitochondrial pool of p66Shc. Methods: The p66Shc and mature HtrA2 cDNA were cloned into eukaryote expression vector, then the constructs were cotransfected into HEK293T cell line, analyze p66Shc by Western blot; Purify proteins by pET System, then conduct in vitro cleavage assay, SDS-PAGE and Coomassie brilliant blue staining; analyze mitochondrial p66Shc protein level in mammalian cells overexpressed full length HtrA2 or knocked down HtrA2; Analyze p66Shc protein level in mitochondria isolated from mnd2 mouse brain. Results: p66Shc was cleaved by HtrA2 in vivo and in vitro; in mammalian cells, overexpressing full length HtrA2 down regulate mitochondrial p66Shc, and knocking down HtrA2 up regulate mitochondrial p66Shc; In mnd2 mouse brain, the mitochondrial p66Shc protein level was higher than that of the wild type mouse(P<0.05). Conclusion: Omi/HtrA2 cleaves p66Shc, and regulates the protein level of mitochondrial p66Shc,suggesting a possible new mechanism of how Omi/HtrA2 protects neuronal cells.
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Received: 17 April 2012
Published: 25 September 2012
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