| Project/Area Number |
19500313
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Nerve anatomy/Neuropathology
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| Research Institution | Hyogo College of Medicine |
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Principal Investigator |
FUJIWARA Noriko Hyogo College of Medicine, 医学部, 准教授 (10368532)
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| Co-Investigator(Kenkyū-buntansha) |
SUZUKI Keiichiro 兵庫医科大学, 医学部, 教授 (70221322)
SAKIYAMA Haruhiko 兵庫医科大学, 医学部, 助教 (30508958)
EGUCHI Hironobu 兵庫医科大学, 医学部, 助教 (60351798)
OOKAWARA Tomomi 兵庫医療大学, 薬学部, 准教授 (50330452)
YOKOE Syunichi 大阪医科大学, 医学部, 助教 (40454756)
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| Project Period (FY) |
2007 – 2009
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| Project Status |
Completed (Fiscal Year 2009)
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| Budget Amount *help |
¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000)
Fiscal Year 2009: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2008: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
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| Keywords | スーパーオキシドジスムターゼ / Cu / Zn-SOD / 抗体 / 筋萎縮性側索硬化症(ALS) / 酸化的翻訳後修飾 / MALDI-TOF-MS / 筋萎縮性側索硬化症 / ALS / 酸化 / 構造変化 / チャージアイソマー |
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| Research Abstract |
Copper-zinc superoxide dismutase (SOD1) plays a protective role against oxidative stress. On the other hand, recent studies suggest that SOD1 itself is a major target of oxidative damage and has its own pathogenicity in various neurodegenerative diseases, including familial amyotrophic lateral sclerosis (FALS). Only human and great ape SOD1s among mammals have the highly reactive free cysteine residue, Cys111, at the surface of the SOD1 molecule. The purpose of this study was to investigate the role of Cys111 in the oxidative damage of the SOD1 protein, by comparing the oxidative susceptibility of recombinant human SOD1 modified with 2-mercaptoethanol at Cys111 (2-ME-SOD1) to wild-type SOD1. Wild-type SOD1 was more sensitive to oxidation by hydrogen peroxide generating fragments and oligomers compared with 2-ME-SOD1. Moreover, wild-type SOD1, but not 2-ME-SOD1, generated an upper shifted band in reducing SDS-PAGE even by air oxidation. Using mass spectrometry and limited proteolysis, this upper band was identified as an oxidized subunit of SOD1 ; the sulfhydryl group (Cys-SH) of Cys111 was selectively oxidized to cysteine sulfinic acid (Cys-SO_2H) and to cysteine sulfonic acid (Cys-SO_3H). The antibody raised against a synthesized peptide containing Cys111-SO_3H reacted with only the Cys111-peroxidized SOD1 by Western blot analysis, and labeled Lewy-body-like hyaline inclusions and vacuole rims in the spinal cord of human SOD1-mutated ALS mice by immunohistochemical analysis. These results suggest that Cys111 is a primary target for oxidative modification and plays an important role in oxidative damage to human SOD1 including FALS mutants.
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