Structural studies of immune responses.

• Project/Area Number: 25440031
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: Kumamoto University
• Principal Investigator: Ikemizu Shinji 熊本大学, 生命科学研究部, 准教授 (60333522)
• Project Period (FY): 2013-04-01 – 2016-03-31
• Project Status: Completed (Fiscal Year 2015)
• Budget Amount: ¥5,200,000 (Direct Cost: ¥4,000,000、Indirect Cost: ¥1,200,000); Fiscal Year 2015: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2014: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2013: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 構造生物学

Outline of Final Research Achievements

Both of interleukin (IL) -27 and IL-23 are heterodimeric cytokines belong to IL-12 family. IL-27 consists of two subunits, p28 and Epstein-Barr virus-induced gene 3 (EBI3), binds two receptors, specific WSX-1 and gp130, a common receptor subunit shared by IL-6 family. IL-27 promotes the differentiation of type 1 T helper (Th1) cells from naive helper T cell cells. IL-27 was expressed on mammalian cells, then purified. Purified IL-27 was able to bind to specific receptor WSX-1.
IL-23 receptors consist of two subunits, specific IL-23R and IL-12Rβ1 which shares with IL-12. IL-12Rβ1 was expressed on insect cells. The purified IL-12Rβ1 was mixed with IL-23, then purified as complexes with gel chromatography. Crystallization trials of the complex have been carried out. IL-23R expression trials was carried out using E. coli, insect cells and mammalian cells. We obtained IL-23R only using mammalian cells with very low yield. IL-23R might need to coexpress with the ligand and IL-12Rβ1.

Research Products

• [Journal Article] Autosomal dominant immune dysregulation syndrome in humans with CTLA-4 mutations. (2014)
  • Author(s): Schubert D, et al.
  • Journal Title: Nature Med. Volume: 20 Issue: 12 Pages: 1410-1416
  • DOI: 10.1038/nm.3746
  • Peer Reviewed
• [Journal Article] Mutants of lymphotoxin-<alpha> with augmented cytotoxic activity via TNFR1 for use in cancer therapy. (2013)
  • Author(s): Morishige, T., Narimatsu, S., Ikemizu, S., Tsunooda, S., Tsutsumi Y., Mukai, Y., Okada, N., Nakagawa, S.
  • Journal Title: Cytokine Volume: 61 Pages: 578-584
  • Peer Reviewed
• [Journal Article] Crystallization and preliminary X-ray analysis of human MTH1 with a homogeneous N-terminus. (2013)
  • Author(s): Koga Y, Inazato M, Nakamura T, Hashikawa C, Chirifu M, Michi A, Yamashita T, Toma S, Kuniyasu A, Ikemizu S, Nakabeppu Y, Yamagata Y.
  • Journal Title: Acta Crystallogr Sect F Struct Biol Cryst Commun Volume: 69(Pt 1) Issue: 1 Pages: 45-48
  • DOI: 10.1107/s1744309112048002
  • Peer Reviewed
• [Journal Article] mpact of antibodies against amyloidogenic transthyretin (ATTR) on phenotypes of patients with familial amyloidotic polyneuropathy (FAP) ATTR Valine30Methionine. (2013)
  • Author(s): Obayashi, K., Tasaki, M., Jono, H., Ueda, M., Shinriki, S., Misumi, Y., Yamashita, T., Oshima, T., Nakamura, T., Ikemizu, S., Anan, I., Suhr, O., & Ando, Y.
  • Journal Title: Clin. Chim. Acta Volume: 419 Pages: 127-131
  • Peer Reviewed
• [Journal Article] Structure and interactions of the human programmed cell death 1 receptor. (2013)
  • Author(s): Cheng, X., Veverka, V., Radhakrishnan, A., Waters, L.C., Muskett, F.W., Morgan, S., Huo, J., Yu, C., Evans, E.J., Leslie, A.J., Griffiths, M., Stubberfield, C., Griffin, R., Henry, A.J., Jansson, A., Ladbury, J.E., Ikemizu, S., Carr, M.D., Davis, S.J.
  • Journal Title: J. Biol. Chem. Volume: 288 Pages: 11771-11785
  • Peer Reviewed