2005 Fiscal Year Final Research Report Summary
Development of a novel assay system of asymmetric dimethylarginine (ADMA), a risk factor for atherosclerosis.
| Project/Area Number |
15590494
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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 |
Laboratory medicine
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| Research Institution | Okayama Prefectural University |
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Principal Investigator |
KIMOTO Masumi Okayama Prefectural University, Fac.Health and Welfare Science, Professor, 保健福祉学部, 教授 (40108866)
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| Co-Investigator(Kenkyū-buntansha) |
TSUJI Hideaki Okayama Prefectural University, Fac.Health and Welfare Science, Professor, 保健福祉学部, 教授 (20093875)
YAMASHITA Hiromi Okayama Prefectural University, Fac.Health and Welfare Science, Assistant Professor, 保健福祉学部, 講師 (70254563)
HIEMORI Miki Okayama Prefectural University, Fac.Health and Welfare Science, Assistant Researcher, 保健福祉学部, 助手 (80326412)
MATSUOKA Hidehiro Kurume University School of Medicine, Department of Internal medicine, Assistant Professor, 医学部, 講師 (80248393)
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| Project Period (FY) |
2003 – 2005
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| Keywords | Nitric oxide / asymmetric dimethylarginine / N^G,N^G-dimethylarginine dimethylaminohydrolase / monoclonal antibody / atherosclerosis / inhibition ELISA / risk factor for cardiovascular disease |
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| Research Abstract |
Asymmetric dimethylarginine (ADMA) is an endogenous competitive inhibitor of nitric oxide synthase (NOS). Elevated ADMA plasma levels have been reported in connection with diseases associated with an impaired endothelial arginine/NO pathway and endothelial dysfunction, such as atherosclerosis, hypercholesterolemia, chronic heart failure, diabetes mellitus, and hypertension. Moreover, ADMA has recently been shown to be a risk factor for cardiovascular diseases. Therefore, there is increasing interest in measuring ADMA levels in human subjects, experimental animals, and cell culture. So far, the quantitative determination of ADMA by high-performance liquid chromatography (HPLC) analysis has been the most widely applied method. We designed specific two assay systems, the first is a method using enzymatic reaction with N^G,N^G-dimethylarginine dimethylaminohydrolase (E.C. 3.5.3.18 : DDAH) that degrades ADMA to citrulline, and the second an enzyme-linked immunosorbent assay (ELISA) using a monoclonal antibody against ADMA. In general, the analytical performance of ELISA is assessed above all by evaluating its specificity, analytical sensitivity, and facility. Therefore, we investigated the development of a novel ELISA assay for ADMA focusing on the second system described above. First we attempted to prepare a monoclonal antibody (mAb) against ADMA using BALB/c mouse immunized with ADMA-MBS-BSA conjugate. The mAb 3C12 obtained reacted specifically ADMA within a variety of amino acids including arginine derivatives and more sensitively to MBS-acylated ADMA than ADMA. We constructed a standard assay system due to the inhibition ELISA using the mAb. The limit of quantitation was 0.4nM, and the mean recovery was 95% after all treatments for the assay system. We determined ADMA levels in human plasma and found good correlation of the values measured by HPLC (r=0.98,p<0.01).
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Research Products
(6 results)
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[Journal Article] Overexpression of dimethylarginine dimethylaminohydrolase (DDAH) reduces tissue ADMA level and enhances angiogenesis.2005
Author(s)
Jacobi, J., Sydow, K., Degenfeld, G., Zhang, Y., Dayoub, H., Wang, B.Y., Patterson, A.J., Kimoto, M., Blau, H.M., Cooke, J.P.
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Journal Title
Circulation, 11(11)
Pages: 1431-1438
Description
「研究成果報告書概要(和文)」より
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[Journal Article] ADMA regulates angiogenesis : genetic and metabolic evidence.2005
Author(s)
Achan, V., HO, H.K., Heeschen, C., Stuehlinger, M., Jang, J.J., Kimoto, M., Vallance, P., Cooke, J.P.
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Journal Title
Vas.Med., 10(1)
Pages: 7-14
Description
「研究成果報告書概要(和文)」より
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[Journal Article] Dimethylarginined dimethylaminohydrolase and endotherial dysfunction in the failing heart.2005
Author(s)
Chen, Y., Li, Y., Zhang, P., Traverse, J.H., Hou, M., Xu, X., Kimoto, M., Bache, R.J.
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Journal Title
Am.J.Physiol.Heart Circ.Physiol., 289(5)
Pages: H2212-H2219
Description
「研究成果報告書概要(和文)」より
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