| Project/Area Number |
08670532
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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 |
内科学一般
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| Research Institution | Kitasato University |
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Principal Investigator |
OHTSUKI Kenzo Professor, Dept.of Molecular Biochem.School of Allied Health Sciences, Kitasato University, 医療衛生学部, 教授 (60124559)
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| Co-Investigator(Kenkyū-buntansha) |
KAWAGUCHI Kiichiro Lecture, Dept.of Pharmacology, School of Pharmaceutical Sciences, Kitasato Unive, 薬学部, 講師 (10146334)
TANAKA Yuetsu Associate Professor, Det.of Immunology, School of Sciences, Kitasato University, 理学部, 助教授 (30163588)
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| Project Period (FY) |
1996 – 1997
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| Project Status |
Completed (Fiscal Year 1997)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 1997: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1996: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Molecular mechanism / Anti-viral effect / Anti-inflammatory effect / Natural substance / Specific receptor / Protein kinase / Signal transduction / Gene expression / 天然薬物 / 特異受容体 / 抗炎症作用 / 抗ウイルス作用 / プロテインキナーゼ |
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| Research Abstract |
The project study was done to investigate (i) purification of the specific binding proteins (gbPs) of anti-inflammatory egent, glycyrrhizin, from GL-sensitive mammalian cells, (ii) identification of gbPs by determination of their N-terminal partial amino acid sequences, and (iii) kinetics of the GL-induced selective inhibition of their physiological activities in vitro.
To purify the gbPs from GL-sensitive mammalian cells or mouse tissues (liver and spleen) , a GL-affinity column was prepared. By using this GL-affinity column, several gbPs were purified to homogenous polypeptides after partial purification of the 1.5 M NaC1 clude extracts by combination with CM-cellulose column cromatography and gel filtration on a Superdex 200pg column (HPLC) . Determination of their N-termina partial amino acid sequences revealed that (i) arachidonate cascade enzymes, such as phospholipase A_2 (PLA_2) , 5'-1ipoxygenase (5'-LOX) and lipocortin I,are identified as gbPs ; (ii) serine protease (p32) , hyaluronidase (p55) comperiment C3 and immunoglobulin (Igg) are identified as gbPs in human serum ; and (iii) 14 kDa secetory PLA_2 (14 kDa sPLA_2 ) and serine protease are identified as gbPs in the synovial fluids of patients with rheumatonoid arthritis. Furthermore, kenitic studies revealed that GL,but not GA,at low levels (1-5muM) inhibits the-physiological activities of these gbPs and also selectively inhibits the CK-II-mediated activation of these gbPs in vitro.
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