| Project/Area Number |
61480443
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Shinshu University School of Medicine (SUSM) |
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Principal Investigator |
KANAI Masamitsu SUSM,Professor, 医学部, 教授 (20020844)
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| Co-Investigator(Kenkyū-buntansha) |
KUMAGAI Toshiko SUSM,Associate, 医学部, 助手 (00020909)
KATO Michimasa SUSM,Associato, 医学部, 助手 (70135143)
KATSUYAMA Tsutomu SUSM,Associate Professor, 医学部, 助教授 (90020809)
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| Project Period (FY) |
1986 – 1987
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| Project Status |
Completed (Fiscal Year 1987)
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| Budget Amount *help |
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1987: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1986: ¥6,000,000 (Direct Cost: ¥6,000,000)
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| Keywords | Retinoid / RBP / CRBP / CRABP / liver / placenta / anti-CRBP / Radioimmunoassay / liver tumor / colon tumor / CRBP(F) / novel CRABP / Fab'-HRP conjugate / RBP assay / subcellular localization / synthesis |
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| Research Abstract |
Vitamin A and its delivatives ( retinoids ) influence a variety of biological process other than vision and reproduction. Serum retinol-binding protein (RBP) is engaged in retinol transport in the blood form liver to target tissues. Within the cell, cellular retinol-binding protein, CRBP and cellular retinoic acid-binding protein, CRABP play important roles for intracellular transport of retinol or retinoic acid and intimately related with the molecular mechanism of retinoid action. In a human study to investigate the distribution of the proteins in various tissues and determine target cells of retinoid action, the following results were obtained.
1. Purification and partial characterization of CRBP and CRABP from human placenta and CRBP from liver were performed with good yields. The proteins exibited similar physicochemical properties with rat counterparts.
2. Anti-human liver CRBP was raised in turkeys and a sensiive RIA system for human CRBP was developed with good precision.
3. CRBP levels in fuman liver tumors were found to be significantly reduced compared with those in the adjacent tissues, whereas no significant difference of the levels was observed between the colon tumors and the adjacent tissues.
4. With regard to intracellular retinoid-binding protein other than CRBP and CRABP, CRBP (F) from piscine eye and a novel CRABP from epididymis were purified and partially characterized.
5. Anti rat RBP Fab' was conjugated to horseradish peroxidase using maleimide compound. The conjugate was successfuly used for the assay of RBP in biological samples and for subcellular localization of RBP in liver and kidney , major sites of its synthesis and catabolism.
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