| Project/Area Number |
11670728
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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 |
Circulatory organs internal medicine
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| Research Institution | Tokyo Metropolitan Organization for Medical Research |
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Principal Investigator |
YAMAGUCHI Atsumi The Tokyo Metropolitan Institute of Medical Science, Researcher(TIMS), Researcher, 東京都臨床医学総合研究所, 研究員 (70124500)
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| Co-Investigator(Kenkyū-buntansha) |
YAMAGISHI Kimiko TIMS, Researcher, 東京都臨床医学総合研究所, 研究員 (20200602)
AKAMATSU Noriko TIMS, Researcher, 東京都臨床医学総合研究所, 研究員 (30124431)
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| Project Period (FY) |
1999 – 2000
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| Project Status |
Completed (Fiscal Year 2000)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2000: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1999: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Keywords | CD36 / phospholipidliposome / oxidized LDL / anti CD36 monoclonal antibody, GS95 / スカベンジャー受容体 |
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| Research Abstract |
CD36 has binding sites of negatively charged lipid such as oxdized LDL and phospholipid liposome. We made a new anti-CD36 monoclonal antibody, GS95, whose epitope was near commercially available anti-CD36 monoclonal antibodies, OKM5 and FA6-152, but not identical. GS95 inhibited bindings both oxdized LDL and negatively charged phospholipid liposome to CD36, while OKM5 and FA6-152 only inhibited binding of oxdized LDL to CD36.
We made mutant CD36 trandfected cells and defined the binding ablity of GS95 to the various mutant CD36 expressed cells. We considered that the decrease of the binding of GS95 to the mutant CD36 expressed cells was the loss of the binding sites of lipid on CD36. We used two strategies to clarify lipid binding sites on CD36. The first was mouse monoclonal antibody couldn't react to mouse antigen and the second was negatively charged mouse chimaera CD25cDNA or amono acid substituted CD36cDNA, subcloned into the expression vector and transfected to COS cells.
OKM5 or FA6-152 didn't bind to human-#155-170 mouse chimaera CD36 expressed cells that coincided with previous report that there were epitopes of them. The epitope of GS95 was suspected to be near those of OKM5 and FA6-152, but GS95 bound to human-#155-170 or #170-183 mouse chimaera CD36 expressed cells. The GS95 bound to mutant CD36s whose lysine and arginine were changed to alanine in #162-178 and #228-243 region. The binding of GS95 to a mutant CD36 whose lysine and arginine were substituted by alanune in #403-407 region was slightly decreased that had a possibility the site was the binding site of phospholipid liposome.
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