| Project/Area Number |
03454157
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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 |
Pathological medical chemistry
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| Research Institution | Tokyo Metropolitan Institute of Medical Science |
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Principal Investigator |
NAGAI Yoshitaka Tokyo Metropolitan Inst. Med. Sci., Direct., 所長 (80072974)
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| Co-Investigator(Kenkyū-buntansha) |
OSANAI Taka Tokyo Metropolitan Inst. Med. Sci., Researcher, 生命情報研究部門, 研究員 (60126018)
SANAI Yutaka Tokyo Metropolitan Inst. Med. Sci., Researcher, 生命情報研究部門, 研究員 (40150289)
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| Project Period (FY) |
1991 – 1992
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| Project Status |
Completed (Fiscal Year 1992)
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| Budget Amount *help |
¥6,700,000 (Direct Cost: ¥6,700,000)
Fiscal Year 1992: ¥3,100,000 (Direct Cost: ¥3,100,000)
Fiscal Year 1991: ¥3,600,000 (Direct Cost: ¥3,600,000)
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| Keywords | ganglioside / sialyltransferase / Xenopus oocyte / photo-affinity label / アフリカツメガエル / 発生 / アフィニティ-標識 / 界面活性剤 / ラクトシルセラミド |
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| Research Abstract |
Isolation and characterization of sialyltransferases which synthesize bioactive gangliosides were carried out.
[1] High sensitive and non-radioactive assay method for sialyltransferases: To improve the assay method of ganglioside sialyltransferase activity, we used pryridylamine-labeled oligosaccharides as substrate.These fluorescent substrates were well separated in HPLC and were applicable in sialyltransferase assay. Ganglioside was digested by endoglycoceramidase and released oligosaccharide was labeled by pyridylamine. Sialyltransferase was partially purified from membrane fraction of rat liver Golgi apparatus. Pyridylamine labelled GM3-oligosaccharide was good substrate for GD3 synthase as native GM3 gangliosede. But, Pyridylamine labelled oligosaccharide of CDH was poor substrate for GM3 synthase. Detergent requirment for full activity of GD3 synthase was reduced suggesting that detergent act in part not only as solubilizer of enzyme but also raise availability of glycolipids
[2] P
… More
hoto-affinity labeling of sialyltransferase: Photoactive diazilin derivatives of lactosylceramide and CMP-NeuAc which were the substrate of sialyltransferase (GM3 synthase). These compounds were good substrates and had high affinity to GM3 synthase, respectively. Some molecules were specifically labeled by these compounds and detected by SDS-PAGE autoradiography.
[3] Characterization and biosynthesis of gangliosides in developing Xenopus laevis: Change of ganglioside species during early development of Xenopus laevis were studied. Major ganglioside in oocyte was GalNAc-GM1b. There is two possible pathways for in vivo synthesis of GalNAc-GM1b. UDP-GalNAc : GM1b beta1-3 N-acetylgalactosaminyl transferase activity was only detected in Xenopus membrane fraction. These suggest that GalNAc-GM1b was synthesized via GM1b from gangliotetraosylceramide in Xenopus. Dramatical elevation of level of GalNAc-GM1b and GM3 was observed after fertilization. Several lines of evidence suggest that alpha2-3 sialyltransferase activity was elevated during this stage. Less
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