| Project/Area Number |
02557103
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| Research Category |
Grant-in-Aid for Developmental Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | Showa University |
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Principal Investigator |
TOMITA Motowo Showa Univ. Pharmaceutical Sci. Professor, 薬学部, 教授 (30102370)
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| Co-Investigator(Kenkyū-buntansha) |
FURUICHI Kiyoshi Yamanouchi Phram. LTD. Res. Center Res. Head, 中央研究所, 研究主任
TAKEMOTO Toshiyuki Yamanouchi Phram. LTD. Res. Center Res. Head, 中央研究所, 研究主任
NAKANO Yasuko Showa Univ. Pharmaceutical Sci. Res. Assist., 薬学部, 助手 (20155790)
MIURA Namho Showa Univ. Pharmaceutical Sci. Res. Assist., 薬学部, 助手 (10146904)
TOBE Takashi Showa Univ. Pharmaceutical Sci. Lecturer, 薬学部, 講師 (90102368)
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| Project Period (FY) |
1990 – 1991
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| Project Status |
Completed (Fiscal Year 1991)
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| Budget Amount *help |
¥14,000,000 (Direct Cost: ¥14,000,000)
Fiscal Year 1991: ¥3,800,000 (Direct Cost: ¥3,800,000)
Fiscal Year 1990: ¥10,200,000 (Direct Cost: ¥10,200,000)
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| Keywords | MACIF / CD59 / regulatory factor of complement / GPl-anchored membrane protein / urine MACIF / 組換え型MACIF / GPI型膜タンパク質 / 発現ベクタ- |
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| Research Abstract |
We had only a limited line of evidence on MACIF when this research had started two years ago. The properties of MACIF we knew at the beginning were that MACIF was a glycosylphosphatidylinositol(GPl)-anchored protein which specifically inhibited the formation of membrane attack complex of homologous complement.
New findings obtained from this reseach project are as follows. (1) it was almost impossible to prepare a large amount of the membrane form of MACIFby using presently available techniques. (2) we tried to prepare the soluble forms of MACIF, instead of the membrane form by three dimerent sauces. The first one was from human urine which contained about 0.5 mg/L of the soluble form. The second and third ones were from Eschericia coli and CHO cells, respectively, in which recombinant soluble forms of MACIF were produced by gene engineering. (3) MACIF consists of 77 amino acids, a N-glycosidic oligosaccharide unit and a GPl-oligosaccharide unit. The urine form had both units, and the recombinant form from E. coli had no units while that from CHO cells had only a N-glycosidic oligosaccharide unit. (4) activities of the three soluble forms were only 0.1% as much as that of the membrane form wheer those activities were assayed by the inhibition of hemolysis using guinea pig erythrocytes and human complement. This result indicated that the membrane binding of MACIF was important for the activity, and the carbohydrate moieties were not essential for the activity. (5) the urine MACIF significantly protected the cytotoxicity caused by recirculation after kidney hypaemia of marmosets, although the reaction mechanism was not clear.
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