| Project/Area Number |
01304058
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| Research Category |
Grant-in-Aid for Co-operative Research (A)
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| Allocation Type | Single-year Grants |
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| Research Field |
代謝生物化学
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| Research Institution | Kyushu University |
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Principal Investigator |
KATO Keitaro Kyushu University, Faculty of Pharmaceuticl Sciences, Professor., 薬学部, 教授 (70037571)
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| Co-Investigator(Kenkyū-buntansha) |
橋本 隆 信州大学, 医学部, 教授 (80009935)
TSUBOI Shozo Yamagata University, School of Medicine, Professor., 医学部, 教授 (70004554)
田代 裕 関西医大, 医学部, 教授 (40077558)
大村 恒雄 九州大学大学院, 医学研究科, 教授 (80029933)
IKEHARA Yukio Fukuoka University, School of Medicine, Professor., 医学部, 教授 (70037612)
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| Project Period (FY) |
1989 – 1990
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| Project Status |
Completed (Fiscal Year 1990)
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| Budget Amount *help |
¥11,100,000 (Direct Cost: ¥11,100,000)
Fiscal Year 1990: ¥5,600,000 (Direct Cost: ¥5,600,000)
Fiscal Year 1989: ¥5,500,000 (Direct Cost: ¥5,500,000)
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| Keywords | Cell organelle / Molecular recognition / Mitochondrion / Lysosome / Peroxisame / Endoplasmic reticulum / Endosome / Nucleus / 細胞オルガネラ / 分子認識 / 形質膜 / エンドサイト-シス / 選別シグナル / リソソ-ム / ゴルジ体 |
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| Research Abstract |
1. A cytosolic factor which plays important roles in the import process of mitochondrial proteins, and the receptor of mitochondrial precursor proteins were purified with an affinity column chromatography using the presequence as a ligand. A precursor processing protease was purified to homogeneity from rat liver mitochondria and its properties were examined. 2. Full length cDNAs for three lysosomal membrane glycoproteins in rat liver were isolated and sequenced, and biosynthesis, processing and intracellular transport of these glycoproteins were studied using pulsechase experiments with primary cultured rat hepatocytes. 3. The structural requirements for the signal-anchor and stop-transfer sequences for the membrane of ER were examined using an in vitro protein translocation system. 4.70kDa peroxisomal membrane protein had the ATP-binding domain which was similar to those of a superfamily involved in membrane transport. On the treatment of isolated rat liver peroxidase with proteinase
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K, this domain (exposed to the cytosol) was cleaved off and acyl-CoA oxidase import was diminished. 5. Using two steps affinity chromatography, anti-DDDED-Sepharose and nucleoplasmin-Sepharose, a protein of 69kDa was purified from nuclear pore fraction and this protein reacted with nuclear transport signal sequences. 6. The primary structure of rat liver 5' -nucleotidase was deduced from the cDNA sequence. This enzyme was initially synthesized as a precursor, cotranslationally processed to an intermediate form containing a hydrophobic domain at the COOH terminus, and finally converted to a mature form by removal of the hydrophobic domain and by simultaneous replacement with glycosyl-phosphatidylinositol. 7. The subpopulation of endosomes was isolated by the high magnetic field separation and the density gradient centrifugation, and effect of microtubles on endocytosis of transferin was studied. 8. SARI, a yeast gene which was originally isolated as a multicopy suppressor of the sec12 temperature-sensitive mutation, encodes a 21kDa GTP-biding protein and is essential for ER-to-Golgi protein transport. Its product, Sarip, is a peripheral membrane protein mainly residing in the ER. Less
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