2006 Fiscal Year Final Research Report Summary
Preparation of the glyco-chip by using endo-type glucan hydrolases and the monitoring of the interaction between biomolecules
| Project/Area Number |
17550150
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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 |
Chemistry related to living body
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| Research Institution | Tokyo Institute of Technology |
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Principal Investigator |
MORI Toshiaki INSTITUTE OF TECHNOLOGY, DEPARTMENT OF BIOMOLECULAR ENGINEERING, ASSOCIATE PROFESSOR, 大学院生命理工学研究科, 助教授 (50262308)
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| Co-Investigator(Kenkyū-buntansha) |
FURUSAWA Hiroyuki INSTITUTE OF TECHNOLOGY, DEPARTMENT OF BIOMOLECULAR ENGINEERING, ASSISTANT PROFESSOR, 大学院生命理工学研究科, 助手 (60345395)
KAWASAKI Takayoshi INSTITUTE OF TECHNOLOGY, DEPARTMENT OF BIOMOLECULAR ENGINEERING, ASSISTANT PROFESSOR, 大学院生命理工学研究科, 助手 (60334504)
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| Project Period (FY) |
2005 – 2006
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| Keywords | glyco-microarray / transglycosylation / endoglycosidase / endo-M / glycomics / N-glycan / glycoconjugate / IPN |
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| Research Abstract |
Glycoconjugates such as glycolipids and glycoproteins on cell membranes play an important role of receptor to lectins, toxins and viruses in living systems, and are related to the formation of organized systems such as cell adhesion, differentiation and morphogenesis. In order to elucidate the function of glycoconjugates on the cell surface, it is necessary to design a suitable glyco-microarray.
We employed two methods to modify the surface of polymer substrates at the level of nm scale using supercritical fluids. The first method is soaking polysaccharides such as hyaluronan and dextran in the scF-swollen polymer substrates. The second method is polymerization such as dextran formation by dextransucrase in the scF-swelled polymer substrates. The hyaluronan immobilized surface was recognized by hyaluronan binding protein. It will be applicable to prepare glycoconjugate-coated polymer materials
Endo-beta-N-Acetylglucosaminidase from Mucor hiemalis (Endo-M) has been shown to have transglycosylation activity as well as hydrolytic activity. We employed two methods to immobilize of N-glycans to the solid surface. One is the tranglycosylations of N-glycopeptide to the GlcNAc bearing a biotin moiety in the buffer solution, and incorporation of the biotinylated N-glycans to the surface covered with Avidins. The other is the direct immobilization of N-glycopeptide to the surface covered with GlcNAc molecules using tranglycosylation by Endo-M. As the Endo-M can be functionalized to not only the high-mannose type and hybrid type of asparagines-linked oligosaccharides but also the complex-type of biantennary oligosacchrides in glycoproteins, we believe that this method will be useful to prepare the various glyco-chip array.
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