2000 Fiscal Year Final Research Report Summary
Synthesis of photo-affinity oligosaccharide probe and LacdiNAc sequence specific for secretory glycoproteins in MDCK cells
| Project/Area Number |
11680618
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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 | SASAKI INSTITUTE |
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Principal Investigator |
OHKURA Takashi SASAKI INSTITUTE, Dept.of Biochemistry, Research, 生化学部, 主任研究員 (50183223)
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| Co-Investigator(Kenkyū-buntansha) |
KUGE Sayuri SASAKI INSTITUTE, Dept.of Biochemistry, Researcher, 生化学部, 研究員 (50260104)
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| Project Period (FY) |
1999 – 2000
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| Keywords | N-linked sugar chain / MDCK cell / lectin / N-acetylalactosamine / secretion / photoaffinity labeling |
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| Research Abstract |
As the first step to identify new intracellular lectins binding to complex type of glycans, photo-affinity oligosaccharide probe was synthesized using sulfo-SBDE.Then, to determine the target glycan for photo-affinity probe, secretory and surface glycoproteins of apical and basolateral sides were separated from metabolically labeled MDCK cells ([^3H]-glucosamine), and the respective N-linked glycans were analyzed. The [^3H]-oligosaccharides released from secretory and surface glycoproteins of apical side had about twice radioactivity than those of basolateral side, but their structures of both sides were equal to each other. Their predominant structures consisted of high mannose type glycans and biantennary complex type glycans with or without bisecting GlcNAc. The most interesting carbohydrate features were LacdiNAc sequences on the non-reducing terminal. Over 20% of oligosaccharides derived from secretory glycoproteins of both sides contained LacdiNAc, in contrast oligosaccharides from both surface glycoproteins did not contain this moiety. Moreover, the LacdiNAc could not be observed in oligosaccharides containing bisecting GlcNAc. These results indicate that the LacdiNAc is synthesized under strict substrate specificities of various glycosyl-transferases and this moiety might play an important role for secretory pathway of MDCK cells.
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