Investigation into the LINE transposition process by structural study of DNA-protein complexes

• Project/Area Number: 23570139
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Multi-year Fund
• Section: 一般
• Research Field: Structural biochemistry
• Research Institution: The University of Tokushima
• Principal Investigator: MAITA Nobuo 徳島大学, 疾患酵素学研究センター, 准教授 (00404046)
• Project Period (FY): 2011 – 2013
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥5,460,000 (Direct Cost: ¥4,200,000、Indirect Cost: ¥1,260,000); Fiscal Year 2013: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000); Fiscal Year 2012: ¥1,820,000 (Direct Cost: ¥1,400,000、Indirect Cost: ¥420,000); Fiscal Year 2011: ¥2,080,000 (Direct Cost: ¥1,600,000、Indirect Cost: ¥480,000)
• Keywords: Ｘ線結晶構造解析 / レトロトランスポゾン / 結晶構造解析

Research Abstract

The aim of this project is to reveal the structural basis of the LINE. To obtain the stable protein, I tested four EN-Myb constructs, but none could be expressed or purified better than Sart1-ORF2p. Next, I used Sf9 cells as protein expression system to overcome the codon-usage issue. Although I detected the ORF1p expression, ORF2p was not co-expressed. I tried to purify the ORF1p expressed in Sf9 cells, however, ORF1p was recovered in insoluble fraction and failed to purify.

Research Products

• [Journal Article] Structural basis for simultaneous recognition of an O-glycan and its attached peptide of mucin family by immune receptor PILRα. (2014)
  • Author(s): Kuroki K, Wang J, Ose T, Yamaguchi M, Tabata S, Maita N, Nakamura S, Kajikawa M, Kogure A, Satoh T, Arase H, Maenaka K.
  • Journal Title: Proc Natl Acad Sci U S A. Volume: 111(24) Issue: 24 Pages: 8877-8882
  • DOI: 10.1073/pnas.1324105111
  • Peer Reviewed / Open Access
• [Journal Article] Structural and clinical implications of amino acid substitutions in α-L-iduronidase: Insight into the basis of mucopolysaccharidosis type I (2014)
  • Author(s): Seiji Saito, Kazuki Ohno, Nobuo Maita, Hitoshi Sakuraba
  • Journal Title: Molecular Genetics and Metabolism Volume: 111 Issue: 2 Pages: 107-112
  • DOI: 10.1016/j.ymgme.2013.10.005
  • Peer Reviewed
• [Journal Article] Observation of the controlled assembly of preclick components in the in situ click chemistry generation of a chitinase inhibitor (2013)
  • Author(s): Hirose T, Maita N, Taniguchi H, Sharpless KB, Omura S, Sunazuka T
  • Journal Title: Proc.Natl.Acad.Sci. USA. Volume: 110 Issue: 40 Pages: 15892-15897
  • DOI: 10.1073/pnas.1315049110
  • Peer Reviewed
• [Journal Article] Human α-L-iduronidase uses its own N-glycan as a substrate-binding and catalytic module (2013)
  • Author(s): Maita N, Tsukimura T, Taniguchi T, Saito S, Ohno K, Taniguchi H, Sakuraba H
  • Journal Title: Proc. Natl. Acad. Sci. USA. Volume: 110 Issue: 36 Pages: 14628-14633
  • DOI: 10.1073/pnas.1306939110
  • Peer Reviewed
• [Journal Article] Crystallographic and NMR evidence for flexibility in oligosaccharyltransferases and its catalytic significance (2013)
  • Author(s): James Nyirenda, et al.
  • Journal Title: Structure Volume: 21 Issue: 1 Pages: 32-41
  • DOI: 10.1016/j.str.2012.10.011
  • Peer Reviewed
• [Journal Article] Crystallization, X-ray diffraction analysis and SIRAS phasing of human α-L-iduronidase (2012)
  • Author(s): Maita N, Taniguchi H, and Sakuraba H
  • Journal Title: Acta Crystallographica Volume: Section F 68(Pt 11) Issue: 11 Pages: 1363-1366
  • DOI: 10.1107/s1744309112040432
  • Peer Reviewed
• [Journal Article] Crystallization strategy for the glycoprotein-receptor complex between measles virus hemagglutinin and its cellular receptor SLAM (2012)
  • Author(s): Hashiguchi T, Ose T, Kubota M, Maita N, Kamishikiryo J, Maenaka K, Yanagi Y.
  • Journal Title: Prot Pept Lett Volume: 19 Issue: 4 Pages: 468-473
  • DOI: 10.2174/092986612799789314
  • NAID: 120006373252
  • Peer Reviewed
• [Journal Article] Crystallographic snapshots of Tom20-mitochondrial presequence interactions with disulfide-stabilized peptides (2011)
  • Author(s): Saitoh T, Igura M, Miyazaki Y, Ose T, Maita N, Kohda D.
  • Journal Title: Biochemistry Volume: 50 Issue: 24 Pages: 5487-5496
  • DOI: 10.1021/bi200470x
  • Peer Reviewed
• [Presentation] A novel function of the N-glycan revealed by the crystal structure of α-L-iduronidase (2014)
  • Author(s): Nobuo Maita
  • Organizer: 第９回研究所ネットワーク国際シンポジウム
  • Place of Presentation: 大阪大学銀杏会館（大阪府）
• [Presentation] In situ click chemistry誘導型キチナーゼ阻害剤をリードとする高活性誘導体の創製 (2013)
  • Author(s): 廣瀬友靖, 他
  • Organizer: 日本薬学会第133年会
  • Place of Presentation: パシフィコ横浜（神奈川県）
• [Presentation] マクロライド骨格をテンプレートとした新規キチナーゼ阻害剤の創製 (2013)
  • Author(s): 菅原章公, 他
  • Organizer: 日本薬学会第133年会
  • Place of Presentation: パシフィコ横浜（神奈川県）
• [Presentation] ヒトα-L-イズロニダーゼの結晶構造解析 (2012)
  • Author(s): 真板宣夫, 月村考宏, 大野一樹, 谷口寿章, 櫻庭均
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場(福岡県)
  • Year and Date: 2012-12-12
• [Presentation] ヒトα-L-イズロニダーゼの結晶構造解析 (2012)
  • Author(s): 真板宣夫, 他
  • Organizer: 第35回日本分子生物学会年会
  • Place of Presentation: 福岡国際会議場（福岡県）
• [Presentation] Crystal structure of T-H protein complex of the glycine cleavage system (2011)
  • Author(s): Okamura-Ikeda K, Hosaka H, Maita N, Fujiwara K, Yoshizawa AC, Nakagawa A, Taniguchi H.
  • Organizer: XXII Congress and General Assembly of the International Union of Crystallography
  • Place of Presentation: Palacio Municipal de Congresos de Madrid (Madrid, Spain)
  • Year and Date: 2011-08-25
• [Presentation] Crystal structure of humanα-L- iduronidase (2011)
  • Author(s): Maita N, Taniguchi H, Sakuraba H.
  • Organizer: XXII Congress and General Assembly of the International Union of Crystallography
  • Place of Presentation: Palacio Municipal de Congresos de Madrid (Madrid, Spain)
  • Year and Date: 2011-08-24
• [Presentation] Crystal structure of Cbl-b TKB domain in complex with Cblin (Cbl-b inhibitor) (2011)
  • Author(s): Ueji T, Shirakata A, KondohS, Nagano K, Maita A, Maita N, Okumura Y, Nikawa T.
  • Organizer: XXII Congress and General Assembly of the International Union of Crystallography
  • Place of Presentation: Palacio Municipal de Congresos de Madrid (Madrid, Spain)
  • Year and Date: 2011-08-24
• [Presentation] Crystal structure of human alpha-L-iduronidase (2011)
  • Author(s): Nobuo Maita et al.
  • Organizer: XXII Congress and General Assembly of the International Union of Crystallography
  • Place of Presentation: Madrid, Spain
• [Remarks]
  • URL: http://pub2.db.tokushima-u.ac.jp/ERD/person/166156/profile-ja.html