Earticle

초록

영어

High-mannose-type oligosaccharides are enzymatically trimmed in the endoplasmic reticulum, giving rise to various processing intermediates with exposure of specific glycotopes that are recognized by a series of lectins involved in glycoprotein-fate determination in cells. Atomic information of dynamic oligosaccharide conformations is essential for a quantitative understanding of energetics of the carbohydrate-lectin interactions. Although carbohydrate NMR spectroscopy is useful for characterizing such conformational dynamics, it is often hampered by poor spectral resolution and the lack of recombinant technique to produce homogeneous glycoforms. To overcome these difficulties, we have recently developed a methodology for preparation of a homogeneous high-mannose-type oligosaccharide with 13C labeling using genetically engineered yeast strain. We herein successfully extended this method to overexpression of 13C-lebeled Man9GlcNAc2 (M9) using a newly engineered yeast strain with deletion of four genes involved in N-glycan processing. This enabled high-field NMR analyses of 13C-labeld M9 in comparison with its processing product M8B, which lacks the terminal mannose residue ManD2. Long-range NOE data indicated that the outer branches can interact with the core in both glycoforms and such foldback conformations are enhanced upon the removal of ManD2. The observed conformational variabilities might be associated with the lectins and the glycan-trimming enzymes.

저자

• Kotaro Yanagi [ Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Graduate School of Pharmaceutical Science, Nagoya City University ]
• Yukiko Kamiya [ Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Graduate School of Pharmaceutical Science, Nagoya City University ]
• Toshihiko Kitajima [ Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology,Tsukuba Central 6, 1-1-1 Higashi, Tukuba, Ibaraki 305-8566, Japan ]
• Takumi Yamaguchi [ Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Graduate School of Pharmaceutical Science, Nagoya City University ]
• Yasunori Chiba [ Research Center for Medical Glycoscience, National Institute of Advanced Industrial Science and Technology,Tsukuba Central 6, 1-1-1 Higashi, Tukuba, Ibaraki 305-8566, Japan ]
• Koichi Kato [ Institute for Molecular Science and Okazaki Institute for Integrative Bioscience, National Institutes of Natural Sciences, Graduate School of Pharmaceutical Science, Nagoya City University, The Glycoscience Institute, Ochanomizu University, RIKEN, Systems and Structural Biology Center, 1-7-22 Suehiro, Tsurumi, Yokohama 230-0045, Japan, GLYENCE Co., Ltd. ]

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간행물

표지보기 관심저널 등록

• 간행물명

  한국당과학회 학술대회

• 간기

  연간

• 수록기간

  2006~2022

• 십진분류

  KDC 517 DDC 614