| Efficient transfer of sialo-oligosaccharide onto proteins by combined use of a glycosynthase-like mutant of Mucor hiemalis endoglycosidase and synthetic sialo-complex-type sugar oxazoline. | |
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MedLine Citation:
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PMID: 20647032 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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BACKGROUND: An efficient method for synthesizing homogenous glycoproteins is essential for elucidating the structural and functional roles of glycans of glycoproteins. We have focused on the transglycosylation activity of endo-ß-N-acetylglucosaminidase from Mucor hiemalis (Endo-M) as a tool for glycoconjugate syntheses, since it can transfer en bloc the oligosaccharide of not only high-mannose type but also complex-type N-glycan onto various acceptors having an N-acetylglucosamine residue. However, there are two major bottlenecks for its practical application: the low yield of the transglycosylation product and the difficulty to obtain the activated sugar oxazoline substrate, especially the sialo-complex type one. METHODS: We carried out the transglycosylation using a glycosynthase-like N175Q mutant of Endo-M, which was found to possess enhanced transglycosylation activity with sugar oxazoline as a donor substrate, in combination with an easy preparation of the sialo-complex-type sugar oxazoline from natural sialoglycopeptide in egg yolk. RESULTS: Endo-M-N175Q showed efficient transglycosylation toward sialo-complex-type sugar oxazoline onto bioactive peptides and bovine ribonuclease B, and each sialylated compound was obtained in significantly high yield. CONCLUSIONS: Highly efficient and simple chemo-enzymatic syntheses of various sialylated compounds were enabled, by a combination of a simple synthesis of sialo-complex-type sugar oxazoline and the Endo-M-N175Q catalyzed transglycosylation. GENERAL SIGNIFICANCE: Our method would be very useful for a practical synthesis of biologically important glycopeptides and glycoproteins. |
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Authors:
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Midori Umekawa; Takayuki Higashiyama; Yurie Koga; Tomonari Tanaka; Masato Noguchi; Atsushi Kobayashi; Shin-Ichiro Shoda; Wei Huang; Lai-Xi Wang; Hisashi Ashida; Kenji Yamamoto |
Publication Detail:
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Type: Journal Article; Research Support, Non-U.S. Gov't Date: 2010-07-18 |
Journal Detail:
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Title: Biochimica et biophysica acta Volume: 1800 ISSN: 0006-3002 ISO Abbreviation: Biochim. Biophys. Acta Publication Date: 2010 Nov |
Date Detail:
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Created Date: 2010-09-06 Completed Date: 2010-11-09 Revised Date: - |
Medline Journal Info:
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Nlm Unique ID: 0217513 Medline TA: Biochim Biophys Acta Country: Netherlands |
Other Details:
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Languages: eng Pagination: 1203-9 Citation Subset: IM |
Copyright Information:
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Copyright © 2010 Elsevier B.V. All rights reserved. |
Affiliation:
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Graduate School of Biostudies, Kyoto University, Kyoto, Japan. mumekawa@umich.edu |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Carbohydrate Conformation Carbohydrate Sequence Glycopeptides / metabolism Glycosylation Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase / genetics*, metabolism* Molecular Sequence Data Mucor / enzymology*, genetics Mutation / genetics* Oligosaccharides / metabolism* Oxazoles / isolation & purification, metabolism* Sialic Acids / metabolism* |
| Chemical | |
Reg. No./Substance:
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0/Glycopeptides; 0/Oligosaccharides; 0/Oxazoles; 0/Sialic Acids; EC 3.2.1.96/Mannosyl-Glycoprotein Endo-beta-N-Acetylglucosaminidase |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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