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中国科学院 生物物理研究所(Institute of Biophysics, Chinese Academy of Sciences)

研究成果

研究成果に関するニュース

2013年8月

中国科学院生物物理研究所内に設置された日中構造ウイルス学・免疫学連携研究室の松田善衛特任教授のグループは、レトロウイルスの感染初期過程における膜融合のリアルタイム解析を可能にする新規レポータータンパク質を開発、改良してきた(Kondo N et al. JBC, 2010; Ishikawa H et al. PEDS 2013, 発表論文リスト参照)。デュアルスプリットプロテイン(DSP)と名付けられたこのレポーターは、膜融合の進行をGFP、ルシフェラーゼ活性の両者で測定できる。今回ペンシルバニア大学Eisenberg 、Cohen教授らによって、DSPアッセイが単純ヘルペスウイルスの膜融合の解析に応用され 、従来法では測定できなかった膜融合動態が明らかにされた(J Virol, 14 August 2013, 10.1128/JVI.01700-13, http://jvi.asm.org/cgi/content/abstract/JVI.01700-13v1?etoc)。この報告は、DSPアッセイ がレトロウイルス以外のウイルスでも有用な解析ツールであることを示した最初の報告である。

論文発表

  1. Wang H, Li X, Nakane S, Liu S, Ishikawa H, Iwamoto A, Matsuda Z.
    Co-expression of foreign proteins tethered to HIV-1 envelope glycoprotein on the cell surface by introducing an intervening second membrane-spanning domain.
    PLoS ONE 9(5):e96790, 2014.
  2. Atanasiu D, Saw WT, Gallagher JR, Hannah BP, Matsuda Z, Whitbeck JC, Cohen GH, Eisenberg RJ.
    A dual split protein-based fusion assay reveals that mutations to HSV glycoprotein gB alter the kinetics of cell-cell fusion induced by HSV entry glycoproteins.
    J Virol. 2013 Aug 14. [Epub ahead of print]
  3. Ishikawa H, Meng F, Kondo N, Iwamoto A and Matsuda Z.
    Generation of a dual-functional split-reporter protein for monitoring membrane fusion using self-associating split GFP.
    Protein Engineering, Design & Selection (2012) doi: 10.1093/protein/gzs051, first published online: August 30, 2012.
  4. Toda T, Kuwahara K, Kondo N, Matsuda Z, Maeda Y, Maeda K, Sakaguchi N. Dynamic appearance of antigenic epitopes effective for viral neutralization during membrane fusion initiated by interactions between HIV-1 envelope proteins and CD4/CXCR4.
    Immunobiology 217(9): 864-872, 2012.
  5. Long Y, Meng F, Kondo N, Iwamoto A, Matsuda Z.
    Conserved arginine residue in the membrane-spanning domain of HIV-1 gp41 is required for efficient membrane fusion.
    Protein Cell. 2011;2(5):369-76.
  6. Kondo N, Miyauchi K, Matsuda Z.
    Monitoring viral-mediated membrane fusion using fluorescent reporter methods.
    Curr Protoc Cell Biol. 2011;Chapter 26:Unit 26.9.
  7. Yang W, Ding X, Deng J, Lu Y, Matsuda Z, Thiel A, Chen J, Deng H, Qin Z.
    Interferon-gamma negatively regulates Th17-mediated immunopathology during mouse hepatitis virus infection.
    J Mol Med. 2011;89(4):399-409.
  8. Liu S, Kondo N, Long Y, Xiao D, Iwamoto A, Matsuda Z.
    Membrane topology analysis of HIV-1 envelope glycoprotein gp41.
    Retrovirology. 2010;7:100.
  9. Miyauchi K, Curran AR, Long Y, Kondo N, Iwamoto A, Engelman DM, Matsuda Z.
    The membrane-spanning domain of gp41 plays a critical role in intracellular trafficking of the HIV envelope protein.
    Retrovirology. 2010;7:95
  10. Kondo N, Miyauchi K, Meng F, Iwamoto A, Matsuda Z.
    Conformational changes of the HIV-1 envelope protein during membrane fusion are inhibited by the replacement of its membrane-spanning domain.
    J Biol Chem. 2010;285(19):14681-8.
  11. Wang J, Kondo N, Long Y, Iwamoto A, Matsuda Z.
    Monitoring of HIV-1 envelope-mediated membrane fusion using modified split green fluorescent proteins.
    J Virol Methods. 2009;161(2):216-22.
  12. Kondo N, Ebihara A, Ru H, Kuramitsu S, Iwamoto A, Rao Z, Matsuda Z.
    Thermus thermophilus-derived protein tags that aid in preparation of insoluble viral proteins.
    Anal Biochem. 2009;385(2):278-85.
  13. Shimizu S, Urano E, Futahashi Y, Miyauchi K, Isogai M, Matsuda Z, Nohtomi K, Onogi T, Takebe Y, Yamamoto N, Komano J.
    Inhibiting lentiviral replication by HEXIM1, a cellular negative regulator of the CDK9/cyclin T complex.
    AIDS. 2007;21(5):575-82.
  14. Futahashi Y, Komano J, Urano E, Aoki T, Hamatake M, Miyauchi K, Yoshida T, Koyanagi Y, Matsuda Z, Yamamoto N.
    Separate elements are required for ligand-dependent and -independent internalization of metastatic potentiator CXCR4.
    Cancer Sci. 2007;98(3):373-9.
  15. Shimizu S, Urano E, Futahashi Y, Miyauchi K, Isogai M, Matsuda Z, Nohtomi K, Onogi T, Takebe Y, Yamamoto N, Komano J.
    Inhibiting lentiviral replication by HEXIM1, a cellular negative regulator of the CDK9/cyclin T complex.
    AIDS. 2007;21(5):575-82.
  16. Chiba-Mizutani T, Miura H, Matsuda M, Matsuda Z, Yokomaku Y, Miyauchi K, Nishizawa M, Yamamoto N, Sugiura W.
    New T-Cell-Based Lines with Two Luciferases for Accurately Evaluating Susceptibility to HIV-1 Drugs.
    J Clin Microbiol. 2007;45(2):477-87.
  17. Miyauchi K, Komano J, Myint L, Futahashi Y, Urano E, Matsuda Z, Chiba T, Miura H, Sugiura W, Yamamoto N.
    Rapid propagation of low-fitness drug-resistant mutants of human immunodeficiency virus type 1 by a streptococcal metabolite sparsomycin.
    Antivir Chem Chemother. 2006;17(4):167-74.
  18. Miyauchi K, Curran R, Matthews E, Komano J, Hoshino T, Engelman DM, Matsuda Z
    Mutations of Conserved Glycine Residues within the Membrane-Spanning Domain of Human Immunodeficiency Virus type 1 gp41 Can Inhibit Membrane Fusion and Incorporation of Env onto Virions.
    Jpn J Infect Dis. 2006;59,77-84.
  19. Miyauchi K, Komano J, Yokomaku Y, Sugiura W, Yamamoto N and Matsuda Z.
    Role of the specific amino acid sequence of the membrane-spanning domain of human immunodeficiency virus type 1 in membrane fusion
    J. Virol. 2005;79, 4720-29.

学会発表

  1. Analysis of protein-protein interaction - application of DSP (Dual Split Protein) system to bacteria
    Dehua Liu, Fanxia Meng, Aikichi Iwamoto, Zene Matsuda
    The 13th Awaji International Forum on Infection and Immunity
    September 23-26, 2014 Nara Prefectural Public Hall, Nara, Japan (poster)
  2. Labeling of HIV-1 envelope glycoprotein within the gp120 variable loops to monitor intracellular trafficking in live cells
    Shuhei Nakane, Aikichi Iwamoto, and Zene Matsuda
    The 13th Awaji International Forum on Infection and Immunity
    September 23-26, 2014 Nara Prefectural Public Hall, Nara, Japan (poster)
  3. Nakane S, Iwamoto A, and Matsuda Z.
    Visualization of HIV-1 envelope glycoprotein in live cells by labeling it with GFP in the gp120 subunit
    The 28th Annual Symposium of the Protein Society, San Diego, CA, USA, July 27-30, 2014.
  4. Wang H, Li X, Nakane S, Liu S, Ishikawa H, Iwamoto A and Matsuda Z.
    Co-expression of foreign proteins tethered to HIV-1 envelope protein by introducing an intervening second membrane-spanning domain.
    Retroviruses, Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA, May19-24, 2014.
  5. Wang H, Li X, Nakane S, Ishikawa H, Iwamoto A, and Matsuda Z.
    Evaluation of neutralizing scFV against the HIV-1 envelope protein: development of a new expression vector with scFV tethered to Env.
    Asian-African Research Forum on Emerging and Reemerging Infections,
    Sendai, Japan, January 20-22, 2014.
  6. Nakane S, Iwamoto A, and Matsuda Z.
    Study of structure-function relationship of the HIV-1 major vaccine target gp120: V4 and V5 loops are tolerant to insertion of foreign protein.
    Asian-African Research Forum on Emerging and Reemerging Infections,
    Sendai, Japan, January 20-22, 2014.
  7. Matsuda Z.
    Viral envelope/cell fusion assays using Dual-Split Protein (DSP).
    Asian-African Research Forum on Emerging and Reemerging Infections,
    Sendai, Japan, January 20-22, 2014.
  8. Nakane S, Iwamoto A, and Matsuda Z.
    V4 and V5 loops of HIV-1 gp120 are tolerant to insertion of foreign protein
    The 9th China-Japan Joint Laboratory Workshop, November 1, 2013
    Institute of Microbiology, Chinese Academy of Sciences, Beijing, China
  9. Wang H, Nakane S, Li X, Ishikawa H, Iwamoto A, and Matsuda Z.
    Expression of scFvs tethered to HIV-1 envelope protein to test their neutralization activity
    The 35th Annual Meeting of the Molecular Biology Society of Japan. Fukuoka, Japan, Dec 11-14, 2012.
  10. Wang H, Nakane S, Li X, Ishikawa H, Iwamoto A, and Matsuda Z.
    A novel mammalian expression vector for scFvs to test their neutralization activity against HIV-1 envelope protein
    Asian-African Research Forum on Emerging and Reemerging Infections (AARF) 2013. Tokyo, Japan, Jan 23-24, 2013.
  11. Ishikawa H, Meng F, Iwamoto A, and Matsuda Z.
    Designing new split proteins by using GFP as“beta-tweezers”.
    Asian-African Research Forum on Emerging and Reemerging Infections (AARF) 2012. Kobe, Japan, Jan 11-12, 2012.
  12. Ishikawa H, Long Y, Meng F, Kondo N, Iwamoto A, and Matsuda Z.
    Arginine residue in the membrane-spanning domain of HIV-1 Env facilitates membrane fusion.
    Asian-African Research Forum on Emerging and Reemerging Infections (AARF) 2012. Kobe, Japan, Jan 11-12, 2012.
  13. Ishikawa H, Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A, and Matsuda Z.
    New strategy to develop a dual-functional split protein using self-associating split GFP
    第34回日本分子生物学会年会、横浜、2011年12月13-16日
  14. Ishikawa H, Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A, and Matsuda Z.
    New strategy to develop a dual-functional split protein using self-associating split GFP
    The 8th China-Japan Joint Laboratory Workshop, Pathogenesis, Gene Regulation and Signal Transduction 2011, Nov 21, 2011, China
  15. Ishikawa H, Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A and Matsuda Z.
    Designing new split proteins by GFP insertion.
    17th International Biophysics Congress, October 30-November 3, 2011, China
  16. Long Y, Meng F, Kondo N, Iwamoto A, and Matsuda Z.
    A highly conserved arginine residue in the membrane-spanning domain of HIV-1 gp41 is required for an efficient membrane fusion
    IUMS (International Union of Microbiological Societies 2011 congress) Sapporo, Japan Sep 11-16, 2011
  17. Ishikawa H, Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A and Matsuda Z.
    Alteration in the split points and linker sequence improved the activity of novel dual functional reporter for membrane fusion.
    25th Anniversary Symposium of Protein Society, Boston, USA , July 23-27,2011
  18. Long Y, Meng, Kondo N, Iwamoto A and Matsuda Z
    A highly conserved arginine residue in the membrane-spanning domain of HIV-1 gp41 is required for an efficient membrane fusion, Retroviruses,
    Cold Spring Harbor Laboratory, Cold Spring Harbor NY, USA. May 2011.
  19. Liu S, Kondo K, Long Y, Xiao D, Iwamoto A, and Matsuda Z.
    Analysis of membrane topology of gp41 subunit of HIV-1 envelope glycoprotein in prokaryotic and mammalian cells
    BMB2010, Dec 7-10, 2010, Kobe, Japan
  20. Matsuda Z
    Intracellular neutralizing epitopes of gp41? - Analysis of membrane topology of gp41 subunit of HIV-1 envelope glycoprotein
    Program for AIDS panel meeting, Awaji Yumebutai, September 10, 2010, Hyogo, Japan.
  21. Liu S, Kondo N, Long Y, Xiao D, Iwamoto A, and Matsuda Z.
    Determination of topology of HIV-1 envelope protein ,
    AARF2010, Nov 11-12, 2010, Hanoi, Vietnam
  22. Ishikawa H, Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A, and Matsuda Z.
    Novel Dual Functional Reporter for Analysis of Membrane Fusion,
    AARF2010, Nov 11-12, 2010, Hanoi, Vietnam.
  23. Kondo N, Long Y, Miyauchi K, Meng F, Iwamoto A, Matsuda Z.
    Mutations in the membrane-spanning domain of HIV-1 gp41 cause a kinetic defect in the membrane fusion, Retroviruses,
    Cold Spring Harbor Laboratory, May 24-29, 2010, USA.
  24. Kondo N, Liu S, Iwamoto A, Matsuda Z.
    Topological analysis of the gp41 subunit of envelope glycoprotein of human immunodeficiency virus type 1 (HIV-1) in prokaryotic and mammalian membranes
    Membrane Proteins:Structure & Function, CSH Asia conference, May 10-14, 2010, Suzhou, China
  25. 松田善衛
    HIV-1エンベロープタンパク質gp41サブユニットの機能
    23回日本エイズ学会学術集会 2009年11月26-28日、名古屋
  26. 近藤直幸、 Long Y、宮内浩典、岩本愛吉、松田善衛
    新規レポータータンパク質を用いた HIV-1 エンベロープタンパク質による膜融合の簡便なリアルタイム解析法の開発とその変異体解析への応用,
    第57回日本ウイルス学会学術集会 2009年10月25-27日、東京
  27. Long Y, Miyauchi K, Meng F, Iwamoto A, Engelman DM, and Matsuda Z.
    The Sequence in the Membrane-spanning Domain (MSD) of HIV-1 gp41 is Critical for the Intracellular Transport of Envelope Protein
    China-Japan Research Collaboration on Emerging and Reemerging Infections: First Five Years and Future, October 21-22, 2009, Beijing, China
  28. Kondo N, Meng F, Iwamoto A and Matsuda Z.
    Real-time Monitoring of HIV-1 Membrane Fusion Using Dual Split Reporter Proteins
    China-Japan Research Collaboration on Emerging and Reemerging Infections: First Five Years and Future, October 21-22, 2009, Beijing, CHina
  29. Kondo N, Long Y, Miyauchi K, Iwamoto A, and Matsuda Z.
    Real-time and quantitative characterization of HIV-1 envelope glycoprotein-mediated membrane fusion using new split reporter proteins
    The 9th Awaji International Forum on Infection and Immunity, Awaji, September 8-11, 2009
  30. Long Y, Miyauchi K, Iwamoto A, and Matsuda Z.
    The Sequence in the Membrane-spanning Domain (MSD) of HIV-1 gp41 is Critical for the Intracellular Transport of Envelope Protein
    The 9th Awaji International Forum on Infection and Immunity, Awaji, September 8-11, 2009
  31. Kondo N, Long Y, Iwamoto A and Matsuda Z.
    Analysis of the effect of Mutations of the Conserved Arginine Residue Located in the Middle of the Membrane-Spanning Domain of the HIV-1 Envelope Protein Subunit gp41 Young Investigator talk
    The 23rd Protein Society Symposium, July25-29, 2009, Boston, USA.
  32. Matsuda Z.
    A versatile dual functional reporter for real-time monitoring of membrane fusion
    The 2009 Harvard School of Public Health - China 6th Annual HIV/AIDS Workshop“HIV/AIDS Treatment Strategies in Poor Resources Settings”Hainan Island, China, March 1-4, 2009
  33. Kondo N, Cheng YX, Meng M, Wang J, Long Y, Iwamoto A and Matsuda Z.
    Dual split reporter protein to monitor the membrane fusion induced by viral envelope proteins
    AARF 2008 (Asian-African Research Forum on Emerging and Reemerging Infections), Dec 14-16, Sapporo, Japan
  34. Long Y, Miyauchi K, Meng F, Iwamoto A, Engelman DM, and Matsuda Z
    The sequence in the membrane-spanning domain (MSD) of HIV-1 gp41 is a critical determinant for the intracellular protein kinesis of HIV-1 envelope protein
    AARF 2008 (Asian-African Research Forum on Emerging and Reemerging Infections), Dec 14-16, 2008 Sapporo, Japan
  35. Liu S, Kondo N, Xiao D, Meng F, Iwamoto A, and Matsuda Z.
    Analysis of membrane topology of gp41 subunit of HIV-1 envelope protein
    AARF 2008 (Asian-African Research Forum on Emerging and Reemerging Infections), Dec 14-16, 2008, Sapporo, Japan
  36. Wang J,Kondo N, Long Y, Iwamoto A, and Matsuda Z.
    Monitoring of HIV-1 envelope-mediated membrane fusion using engineered split green fluorescent proteins
    AARF 2008 (Asian-African Research Forum on Emerging and Reemerging Infections),Dec 14-16, 2008, Sapporo, Japan
  37. 近藤直幸, 海老原章郎, Ru H, 倉光成紀, 岩本愛吉, 松田善衛
    高度好熱菌 Thermus thermophilus の構造ゲノム解析によって蓄積された情報の利用による、新規タグ蛋白質の開発
    BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会) 、2008年12月9-12日、神戸
  38. 松田善衛, 宮内浩典, Long Y, Liu S, Meng F, 近藤直幸, Engelman DM, 岩本愛吉
    HIV-1 エンベロープ蛋白質 (Env) 膜貫通部分内 GXXXG モチーフと荷電性アミノ酸の相互関係の Env 細胞内分布への影響
    BMB2008(第31回日本分子生物学会年会・第81回日本生化学会大会 合同大会)、2008年12月9-12日、神戸
  39. 松田善衛, Liu S, Xiao D, Meng F, Engelman DM, 近藤直幸 岩本愛吉
    HIV-1 エンベロープ蛋白質サブユニットgp41膜貫通部分の膜内トポロジーの解析
    第56回ウイルス学会学術集会、2008年10月26-28日、岡山
  40. 松田善衛, 宮内浩典, Long Y, Meng F, Engelman DM, 近藤直幸, 岩本愛吉
    HIV-1 エンベロープ蛋白質サブユニットgp41膜貫通部分の変異体解析
    第56回ウイルス学会学術集会、2008年10月26-28日、岡山
  41. Kondo N, Ebihara A, Ru H, Kuramitsu S, Iwamoto A and Matsuda Z
    Development of protein tags with defined structures for expression and solubilization of HIV proteins
    第8回あわじしま 感染症・免疫フォーラム2008 Sep7-11、淡路
  42. Zene Matsuda, Kosuke Miyauchi, Yufei Long, Shujun Liu, Dan Xiao1, Fanxia Meng, Donald M. Engelman, Aikichi Iwamoto
    The relationship between the GXXXG motif and a potentially charged arginine residue within the membrane-spanning domain of HIV-1 gp41 is critical for the biosynthesis of Env
    第8回あわじしま 感染症・免疫フォーラム/日米エイズ部会 2008 Sep7-11,淡路
  43. Kondo N, Ebihara A, Ru H, Kuramitsu S, Iwamoto A, Matsuda Z.
    Use of Thermus thermophilus Proteins as A Fusion Partner to Enhance Expression-Level and Solubility of Target Proteins Derived from Primate Lentiviruses (HIV/SIV)
    The 22nd Protein Society Symposium, July 19-23, 2008, San Diego, USA
  44. Liu S, Xiao D, Meng F, Engelman DM, Iwamoto A and Matsuda Z.
    Probing a membrane topology of HIV-1 gp41 with two prokaryotic systems
    The 7th Japan-China International Conference of Virology, June 1-3, 2008, Tokyo Japan
  45. Kondo N, Ebihara A, Ru H, Kuramitsu S, Iwamoto A and Matsuda Z.
    A new strategy to facilitate structural analysis of highly insoluble viral proteins derived from HIV-1
    1st China-Japan Joint Workshop on New-Generation Vaccines for Infectious Diseases - Molecular Biology of Host-Pathogen Interaction, May 22, 2008, Beijing, PR China
  46. Kondo N, Ru H, Cheng Y, Ebihara A, Kuramitsu S, Iwamoto A and Matsuda Z.
    Structural analyses of lentiviral proteins: Seeking for fusion partners for crystallization.
    ARF 2008, Jan 28-29 , Osaka, Japan
  47. Liu S.
    Analysis of HIV-1 envelope protein - membrane topology and neutralizing epitopes
    International Symposium on Basic and Applied Immunobiology, Oct. 29-30, 2007, Beijing, PR China
  48. Kondo N.
    Determination of structures of viral proteins of HIV and SIV - search for new tag proteins for crystallization,
    International Symposium on Basic and Applied Immunobiology, Oct. 29-30, 2007, Beijing, PR China
  49. Matsuda Z.
    Role of the membrane-spanning domain (MSD) of HIV-1 envelope (Env) protein in HIV-1 life cycle
    China-Japan Joint-lab Symposium on Emerging Pathogens and Immunology 2007, August 11, 2007, Beijing, China
  50. Matsuda Z, Miyauchi K, Curran R, Matthews E, Komano J, Yamamoto N, and Engelman DM
    The alteration of the phase in the membrane-spanning helix of HIV-1 gp41 negatively affects intracellular transport of HIV-1 envelope protein
    The 21st Protein Society Symposium, July 21-25, 2007, Boston, USA
  51. Miyauchi K, Curran R, Matthews E, Komano J, Murakami T, YamamotoN, Engelman DM and Matsuda Z
    The Specific Phase of Membrane-spanning Helix of HIV-1 gp41 is Critical for Intracellular Transport of Env
    Retroviruses, Cold Spring Harbor Laboratory , May 19-24, 2007, Cold Spring Harbor NY, USA.
  52. Miyauchi K. Curran R, Komano J, Murakami T, Yamamoto N, Engelman DM and Matsuda Z.
    The rotational phase of the localized regionof gp41 membrane-spanning domain alpha-helix affected the Env biogenesis. Retroviruses,
    Cold Spring Harbor Laboratory , May 24-29, 2005 Cold Spring Harbor NY, USA.
  53. Miyauchi K. Curran R, Komano J, Murakami T, Yamamoto N, Engelman DM and Matsuda Z.
    The role of the additional glycine residue within a conserved GXXXG motif of the membrane-spanning domain of HIV-1 Retroviruses,
    Cold Spring Harbor Laboratory , May 24-29, 2005 Cold Spring Harbor NY, USA.
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