Structural studies on the pathological mechanism of dystroglycanopathy

• Project/Area Number: 15K18496
• Research Category: Grant-in-Aid for Young Scientists (B)
• Allocation Type: Multi-year Fund
• Research Field: Structural biochemistry
• Research Institution: The University of Tokyo (2016); Institute of Physical and Chemical Research (2015)
• Principal Investigator: Nagae Masamichi 東京大学, 大学院薬学系研究科(薬学部), 特任研究員 (60619873)
• Project Period (FY): 2015-04-01 – 2017-03-31
• Project Status: Completed (Fiscal Year 2016)
• Budget Amount: ¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000); Fiscal Year 2016: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000); Fiscal Year 2015: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: 糖鎖生物学 / X線結晶構造解析 / 生化学 / 構造生物化学 / 構造生物学 / Ｘ線結晶構造解析

Outline of Final Research Achievements

Congenital muscular dystrophy is a group of diseases caused by defects in O-mannose glycosylation of the alpha-subunit of dystroglycan and progressive weakness and wasting of skeletal muscle are commonly observed. Alpha-dystroglycan (a-DG) is a component of the dystrophin-glycoprotein complex of skeletal muscle cells and directly links several other components to the basement membrane. The abnormal O-mannosylation of a-DG leads to severe congenital muscular dystrophies due to detachment of extracellular matrix proteins from the basal membrane. Phosphorylation at C6-position of O-mannose catalyzed by protein O-mannosyl kinase (POMK) is a crucial step in the biosynthetic pathway of O-mannose glycan. In this project, we solved the crystal structures of POMK catalytic domain in the absence and presence of substrates. These structures provides atomic insights into catalytic reaction mechanism and substrate recognition. These results lead to clinical approach to this severe disease.

Research Products

• [Journal Article] 3D structural analysis of protein O-mannosyl kinase, POMK, a causative gene product of dystroglycanopathy (2017)
  • Author(s): M.Nagae, S.K.Mishra, M.Neyazaki, R.Oi, A.Ikeda, N.Matsugaki, S.Akashi, H.Manya, M.Mizuno, H.Yagi, K.Kato, T.Senda, T.Endo, T.Nogi, and Y.Yamaguchi
  • Journal Title: Genes Cells Volume: - Issue: 4 Pages: 348-359
  • DOI: 10.1111/gtc.12480
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Crystallographic analysis of murine p24γ2 Golgi dynamics domain (2017)
  • Author(s): Nagae M, Liebschner D, Yamada Y, Morita-Matsumoto K, Matsugaki N, Senda T, Fujita M, Kinoshita T, Yamaguchi Y
  • Journal Title: Proteins Volume: 85 Issue: 4 Pages: 764-70
  • DOI: 10.1002/prot.25242
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] 3D Structure and Interaction of p24β and p24δ Golgi Dynamics Domains: Implication for p24 Complex Formation and Cargo Transport. (2016)
  • Author(s): Nagae M, Hirata T, Morita-Matsumoto K, Theiler R, Fujita M, Kinoshita T, Yamaguchi Y.
  • Journal Title: J Mol Biol. Volume: 428 Issue: 20 Pages: 4087-99
  • DOI: 10.1016/j.jmb.2016.08.023
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Enhancement of solubility and yield of a β-glucan receptor Dectin-1 C-type lectin-like domain in Escherichia coli with a solubility-enhancement tag. Protein (2016)
  • Author(s): Dulal HP, Nagae M, Ikeda A, Morita-Matsumoto K, Adachi Y, Ohno N, Yamaguchi Y.
  • Journal Title: Expr Purif. Volume: 123 Pages: 97-104
  • DOI: 10.1016/j.pep.2016.04.002
  • Peer Reviewed / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Crystal structure of human dendritic cell inhibitory receptor (DCIR) C-type lectin domain reveals the binding mode with N-glycan (2016)
  • Author(s): Nagae M, Ikeda A, Hanashima S, Kojima T, Matsumoto N, Yamamoto K, Yamaguchi Y.
  • Journal Title: FEBS Lett. Volume: Mar 26 Issue: 8 Pages: 1-9
  • DOI: 10.1002/1873-3468.12162
  • Peer Reviewed / Open Access / Int'l Joint Research / Acknowledgement Compliant
• [Journal Article] Atomic visualization of a flipped-back conformation of bisected glycans bound to specific lectins (2016)
  • Author(s): Nagae M, Kanagawa M, Morita-Matsumoto K, Hanashima S, Kizuka Y, Taniguchi N, Yamaguchi Y.
  • Journal Title: Sci Rep. Volume: 6 (22973) Issue: 1 Pages: 1-11
  • DOI: 10.1038/srep22973
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Journal Article] Sugar recognition and protein-protein interaction of mammalian lectins conferring diverse functions (2015)
  • Author(s): Nagae M, Yamaguchi Y.
  • Journal Title: Curr Opin Struct Biol. Volume: 34 Pages: 108-15
  • DOI: 10.1016/j.sbi.2015.08.005
  • Peer Reviewed / Acknowledgement Compliant
• [Journal Article] In situ visualization of a glycoform of transferrin: Localization of α2,6-sialylated transferrin in the liver (2015)
  • Author(s): Yuka Matsumoto, Toshie Saito, Kyoka Hoshi, Hiromi Ito, Yoshinobu Kariya, Masamichi Nagae, Yoshiki Yamaguchi, Yoshiaki Hagiwara, Noriaki Kinoshita, Ikuo Wada, Kiyoshi Saito, Takashi Honda and Yasuhiro Hashimoto
  • Journal Title: J. Biochem. Volume: 157(4) Issue: 4 Pages: 211-216
  • DOI: 10.1093/jb/mvu071
  • Peer Reviewed / Open Access / Acknowledgement Compliant
• [Remarks] 折れ曲がった形の糖鎖を可視化 －糖鎖構造の揺らぎの理解へ一歩前進－
  • URL: http://www.riken.jp/pr/press/2016/20160323_2/