| Project/Area Number |
14580625
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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 |
Structural biochemistry
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| Research Institution | Osaka University (2004)
Kyoto Institute of Technology (2002-2003) |
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Principal Investigator |
NATSUKA Shunji Osaka University, Graduate School of Science, Associate Professor, 理学研究科, 助教授 (00243163)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2002: ¥1,600,000 (Direct Cost: ¥1,600,000)
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| Keywords | Vertebrates / Zebrafish / Neurula / Glycoproteins / Glycoconjugates / Lewis antigens / Fucosyltransgerases / 糖蛋白質 / ルイス型抗 |
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| Research Abstract |
We have analyzed expression period and substrate specificity of zebrafish a(1,3)fucosyltransferase (zFT1). As a results, the zFT1 expressed specifically during segmentation period of embryogenesis, and could synthesize Lewis x structure, but not sialyl Lewis x or Lewis a structures. The zFT1 preferred to transfer fucose residue to inner N-acetyllactosamine unit rather than the terminal unit when tandem tri-N-acetyllactosamine oligosaccharide was used as a substrate. The results of in situ hybridization showed that the zFT1 expressed dispersedly at neural tube and around. We have investigated the structure of N-linked glycans expressed in zebrafish embryo. The expression of biantenna-complex-type glycans was remarkably increased from segmentation period. We searched also Lewis x-containing glycans, and found that the biantenna-complex-type glycans had one or two Lewis x structures at their non-reducing end. Those glycans expressed in segmentation period simultaneously with zFI1. We discovered unusual glycan structure that had additional Galb1-4 residue attached to N-acetyllactosamine structure, and also free oligosaccharides with similar structure. As a result of searching for core proteins of Lewis x containing biantenna-complex-type glycans, a glycoprotein with a molecular weight of 62,000 was found as a carrier protein from zebrafish embryos in segmentation period. We designated the glycoprotein as zGP62. Furthermore, we measured the zFT1 activity against Galb1-4Galb1-4GlcNAcb1-R structure, and zFT1 could not utilized that structure as a substrate. Thus it was speculated that the Galb1-4Galb1-4GlcNAcb1-R structure was synthesized by an unknown b1-4galactosyltransferase which could act on Lewis x structure.
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