2016 Fiscal Year Final Research Report
Semi-synthesis of bioactive large glycoproteins
| Project/Area Number |
26248040
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| Research Category |
Grant-in-Aid for Scientific Research (A)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Bio-related chemistry
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| Research Institution | Osaka University |
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Principal Investigator |
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| Project Period (FY) |
2014-04-01 – 2017-03-31
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| Keywords | 糖鎖関連化学 / 糖鎖 / 糖タンパク質 |
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| Outline of Final Research Achievements |
In order to synthesize large bioactive glycoproteins, we examined semisyntheses employing synthesis of glycopeptide by solid phase peptide synthesis (SPPS) and synthesis of long polypeptide in E. Coli expression. Glycopeptide-thioesters bearing human biantennary complex type or highmannose type oligosaccharides were synthesized by Boc-SPPS. Peptide expression in E. Coli was designed to yield either HisTag-Met-target-Cys-peptide or HisTag-ENLYFQ-target-Cys-peptide. These peptides prepared in E. Coli were converted into corresponding Cys-peptides by CNBr treatment or TEV-protease. Native chemical ligation of these peptides and glycopeptide-thioesters yielded Fc domain of human-antibody and sialyltransferase. Folding experiments successfully yielded enzymatically active sialyltransferase. These trials proved that large bioactive glycoprotein consisting over 300 amino acids can be prepared by chemical synthesis.
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| Free Research Field |
生体関連化学
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