Structural basis for N-linked glycosylation site occupancy

2014 Fiscal Year Final Research Report

• Project/Area Number: 24370047
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Partial Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kyushu University
• Principal Investigator: KOHDA Daisuke 九州大学, 生体防御医学研究所, 教授 (80186618)
• Co-Investigator(Renkei-kenkyūsha): SHIMADA Atsushi (70391977) ; MAYANAGI Kouta (50418571)
• Project Period (FY): 2012-04-01 – 2015-03-31
• Keywords: Ｎ型糖鎖修飾 / 翻訳後修飾 / Ｘ線結晶解析 / 膜タンパク質 / 糖転移酵素

Outline of Final Research Achievements

The addition of oligosaccharide chains on the asparagine residues in proteins is catalyzed by the membrane enzyme, oligosaccharyltransferase (OST). We determined the crystal structures of the C-terminal globular domains and the full-length OST protein from hyperthermophilic archaea. We also showed that the dynamic feature of the C-terminal globular domains was essential for the oligosaccharyl transfer activity. The crystal structure in the complex with the oligosaccharide acceptor substrate was determined using the tethering of a peptide containing the Asn-Val-Thr sequon to the protein through a disulfide bond. Toward the determination of the complex structure with the oligosaccharide donor substrate, we analyzed the chemical structures of archaeal lipid-linked oligosaccharides to synthesize the water-soluble analogues.

Free Research Field

構造生物学