| Project/Area Number |
10045076
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| Research Category |
Grant-in-Aid for Scientific Research (A).
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Circulatory organs internal medicine
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| Research Institution | Tokai University |
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Principal Investigator |
NAKAZAWA Hiroe Tokai University, School of Medicine, Professor, 医学部, 教授 (20110885)
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| Co-Investigator(Kenkyū-buntansha) |
ICHIMORI Kouji Tokai Univ., School of Medicine, Assistant Professor, 医学部, 講師 (60184636)
ISHIDA Hideyuki Tokai Univ., School of Medicine, Assistant Professor, 医学部, 講師 (20222424)
ABU・SOUD Hus ケースウエスタンリザーブ大学, 医学部, 主任研究員
IKEDAーSAITO マサオ ケースウエスタンリザーブ大学, 医学部, 教授
STUEHR Denni ケースウエスタンリザーブ大学, 医学部, 教授
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| Project Period (FY) |
1998 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥10,600,000 (Direct Cost: ¥10,600,000)
Fiscal Year 2000: ¥2,300,000 (Direct Cost: ¥2,300,000)
Fiscal Year 1999: ¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 1998: ¥4,400,000 (Direct Cost: ¥4,400,000)
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| Keywords | nitric oxide / nitric oxide synthase / tetrahydrobiopterin (BH4) / arginine / heme-iron / NADPH / テトラバイオプテリン / テトラバイオオプテリン / ヘム |
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| Research Abstract |
Nitric oxide synthase (NOS) transfers electrons supplied with NADPH to oxygen on heme of cytochrome P450-type and oxidizes L-arginine to produce NO during enzyme turnover. But, if the activated oxygen cannot react with L-arginine, the oxygen is released from NOS as reactive oxygen species (ROS). This phenomenon which is called uncoupling reaction, was clarified in detail in each NOS isoform and dependency upon cofactor and substrates was determined. Generations of NO, superoxide (・O_2^-), and H_2O_2 were measured at 25℃ with the metHb formation from HbO_2, the reduction of succinyl cytochrome c, and the generation of Fe (SCN)_4 from Fe^<2+>, respectively. NADPH consumption was also measured to determine total electron flux. Under BH_4^- and arginine-saturated condition, NOS-I and NOS-II did not show significant uncoupling. However, in NOS-III almost a half of electrons remained unconsumed during NO synthesis. In all NOSs, the depletion of substrate, arginine led to uncoupling reaction, whose IC_<50> corresponded to the arginine affinity to each NOS isoform. Under uncoupling condition, major ROS generated from NOS was superoxide in NOS-I and NOS-III, whereas it was H_2O_2 for NOS-II.For NOS-I and NOS-II, the change in electron transfer rate by substrates or cofactors did not alter the ・O_2^-/H_2O_2 ratio, whereas, for NOS-III, the increase in electron transfer rate enhanced ・O_2^- generation. These differences of three NOS isoforms may have an impact on its role in the pathogenesis of NOS-related inflammation processes.
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