Expression of catabolic pathway of asparagine-linked oligosaccharides on plasma membrane and apoptosis
| Project/Area Number |
14580628
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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 City University |
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Principal Investigator |
ITO Kazuo Osaka City University, Graduate School of Science, Associate Professor, 大学院・理学研究科, 助教授 (20183171)
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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 |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2004: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 2003: ¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 2002: ¥2,100,000 (Direct Cost: ¥2,100,000)
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| Keywords | endo-β-N-acetylglucosaminidase HS / endo HS / glycoproteins / asparagine-linked oligosaccharide / エンド-β-N-アセチルグルコサミニダーゼHS / エンドHS / エンド-β-NアセチルグルコサミニダーゼHS |
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| Research Abstract |
1.Verification and specification of target glycoproteins deglycosylated by Endo HS
Target glycoprotein(TGP) deglycosylated by Endo HS was detected from attached epithelial cell in the human oral cavity more than detached cell, but not in leukocyte. The protein of molecular weight(MW) of 55K from the attached epithelial cell was specified as TGP55 by in-gel deglycosylation method. The proteins of MW of 20K, 50K and 105〜110K were also specified as TGP20, TGP50 and TGP105〜110.
2.Identification of TGP55 and TGP50
TGP55 and TGP50 were purified. The N-terminal amino acid was thought to be acetylated. The N-terminal amino acid sequence of deacetylated TGP55 showed high homology with the inner amino acid sequence of human cytokeratin 13, but no proteins having the same N-terminal amino acid sequence was not observed by homology search. The N-terminal amino acid of TGP50 was also blocked. The N-terminal and inner amino acid sequence of TGP50 showed high homology with those of human type I cytokerain 13, indicating that TGP50 is type I cytokeratin 13 or a similar protein to it. Human type I cytokeratin 13 has an asparagine of potential glycosylation site. Endo HS was thought to removing the asparagine-linked oligosaccharide at the site and taking part in the differentiation of the epithelial cell.
3.Immunochemical properties and cellular localization of TGP55 analyzed using monoclonal antibodies against TGP55
Three monoclonal antibodies against TGP55 were prepared. Each antibody reacted with TGP55 and other proteins of attached and detached epithelial cell, the kinds of proteins were different between attached and detached epithelial cell. The outskirts part of the epithelial cell was stained by the antibodies, indicating that TGP55 localizes around the outskirt part of the epithelial cell and the localization changes after peeling of the epithelial cell.
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Report
(4 results)
Research Products
(19 results)
