Folding of cysteine-rich peptides on phospholipid membrane

• Project/Area Number: 24651242
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Single-year Grants
• Research Field: Living organism molecular science
• Research Institution: Osaka University
• Principal Investigator: HARA Toshiaki 大阪大学, 理学(系)研究科(研究院), 特任講師(常勤) (90528233)
• Project Period (FY): 2012-04-01 – 2014-03-31
• Project Status: Completed (Fiscal Year 2013)
• Budget Amount: ¥4,030,000 (Direct Cost: ¥3,100,000、Indirect Cost: ¥930,000); Fiscal Year 2013: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2012: ¥2,600,000 (Direct Cost: ¥2,000,000、Indirect Cost: ¥600,000)
• Keywords: 脂質膜 / ペプチド化学

Research Abstract

In this study, we developed a new protocol for the oxidative folding reaction of cysteine-rich peptides in a phospholipid membrane: "membrane induced folding reaction". We investigated the effect of the phospholipid composition and pH of the reaction on the folding yield and purity of cystein-knot peptides including beta defensins, conotoxins, and other peptides. The best yield was obtained in the presence of glycero-phosholipid membrane with negative charge without adding anr redox reagents. The phospholipid membranes for the folding reaction can be a general method for peptide and protein engineering.

Research Products

• [Journal Article] Folding of cysteine-rich peptides on phospholipid membranes. (2014)
  • Author(s): Hara1, T.; Purwati, E.M.; Wada, A.; Hasegawa, M.; Murata, M.; Aimoto, S.
  • Journal Title: Peptides Science 2013 Volume: 未定
  • Peer Reviewed
• [Journal Article] Purification of Transmembrane Peptides using RP-HPLC. (2014)
  • Author(s): Huang, Y.; Hara, T.; Murata M.
  • Journal Title: Peptides Science 2013 Volume: 未定
  • Peer Reviewed
• [Journal Article] Differential Effects of Peptides Stomagen, EPF1, and EPF2 on Activation of the MAP Kinase MPK6 and the SPCH Protein Level (2013)
  • Author(s): Pawan, K. J., Hara, T., Tanaka, H., Kondo, T., Betsuyaku, S., Sawa, S., Sakagami, Y., Aimoto, S., and Kakimoto, T.
  • Journal Title: Plant Cell Physiol Volume: 54 Pages: 1253-1262
  • NAID: 10031195179
  • Peer Reviewed
• [Journal Article] Structure of the human heart fatty acid-binding protein in complex with the fluorescent probe 1-anilinonaphtalene-8-sulphonic acid (2013)
  • Author(s): Hirose, M., Sugiyama, S., Ishida, H., Niiyama, M., Matsuoka, D., Hara, T., Mizohata, E., Murakami, S., Inoue, T., Matsuoka, S. and Murata, M.
  • Journal Title: Journal of Synchrotron Radiation Volume: 20 Pages: 923-928
  • Peer Reviewed
• [Journal Article] Purification of Transmembrane Peptides using RP-HPLC., Ed. Teshima T. and Nishiushi Y (2013)
  • Author(s): Huang, Y.; Hara, T.; Murata M.
  • Journal Title: Peptides Science 2013 Volume: (in press)
  • Peer Reviewed
• [Journal Article] Folding of cysteine-rich peptides on phospholipid membranes (2013)
  • Author(s): Hara1, T.; Purwati, E.M.; Wada, A.; Hasegawa, M.; Murata, M.; Aimoto, S.
  • Journal Title: Peptides Science 2013 Volume: (in press)
  • Peer Reviewed
• [Journal Article] Structure of the human heart fatty acid-binding protein in complex with the fluorescent probe 1-anilinonaphtalene-8- sulphonic acid. (2013)
  • Author(s): Hirose, M., Sugiyama, S., Ishida, H., Niiyama, M., Matsuoka, D., Hara, T., Mizohata, E., Murakami, S., Inoue, T., Matsuoka, S. and Murata, M.
  • Journal Title: Journal of Synchrotron Radiation Volume: 20 Pages: 923-928
  • Peer Reviewed
• [Journal Article] Differential Effects of Peptides Stomagen, EPF1, and EPF2 on Activation of the MAP Kinase MPK6 and the SPCH Protein Level. (2013)
  • Author(s): Pawan, K. J., Hara, T., Tanaka, H., Kondo, T., Betsuyaku, S., Sawa, S., Sakagami, Y., Aimoto, S., and Kakimoto, T.
  • Journal Title: Plant Cell Physiol. Volume: 54 Pages: 1253-1262
  • NAID: 10031195179
  • Peer Reviewed
• [Presentation] Purification of Transmembrane Peptides using RP-HPLC (2013)
  • Author(s): Huang, Y.; Hara, T.; Murata M.
  • Organizer: 4th Asia-Pacific International Peptide Symposium & 50th Japanese Peptide Symposium
  • Place of Presentation: 大阪(ポスター発表)
• [Presentation] Folding of cysteine-rich peptides on phospholipid membranes (2013)
  • Author(s): Hara, T.; Purwati, E.M.; Wada, A.; Hasegawa, M.; Murata, M.; Aimoto, S.
  • Organizer: 4th Asia-Pacific International Peptide Symposium & 50th Japanese Peptide Symposium
  • Place of Presentation: 大阪(ポスター発表)
• [Presentation] ヒト心筋型脂肪酸結合タンパク質FABP3と蛍光プローブ1-anilinonaphtalene-8-sulphonic acidの複合体結晶構造解析 (2013)
  • Author(s): 廣瀬未果、松岡茂・石田英子・松岳大輔・原利明・溝端栄一・村上聡・井上豪・村田道雄・杉山成
  • Organizer: 日本結晶学会平成25年度年会
  • Place of Presentation: 熊本(ポスター発表)
• [Presentation] Structure of the human muscle fatty acid-binding protein complexed with competitive inhibitor (2013)
  • Author(s): Mika Hirose, Shigeru Sugiyama, Hanako Ishida, Daisuke Matsuoka, Toshiaki Hara, Eiichi Mizohata, Tsuyoshi Inoue, Shigeru Matsuoka, Michio Murata
  • Organizer: The International Conference on Structural Genomics 2013 (ICSG2013)第7回国際構造ゲノム会議
  • Place of Presentation: 北海道(札幌)(ポスター発表)
• [Presentation] Novel fatty acid analogues for the understanding of lipid-protein interactions (2013)
  • Author(s): Hanako Ishida, Shigeru Sugiyama, Mika Hirose, Sebastien Lethu, Hikaru Ano, Fuminori Sato, Daisuke Matsuoka, Toshiaki Hara, Eiichi Mizohata, Tsuyoshi Inoue, Shigeru Matsuoka, Michio Murata
  • Organizer: The International Conference on Structural Genomics 2013 (ICSG2013)第7回国際構造ゲノム会議
  • Place of Presentation: 北海道(札幌)(ポスター発表)
• [Presentation] Structure of the human muscle fatty acid-binding protein complexed with Hydrophobic ligand (2013)
  • Author(s): Shigeru Sugiyama, Hanako Ishida, Daisuke Matsuoka, Toshiaki Hara, Eiichi Mizohata, Tsuyoshi Inoue, Shigeru Matsuoka, Michio Murata
  • Organizer: 4th International Symposium on Diffraction Structural Biology(第4回回折構造生物国際シンポジウム2013)
  • Place of Presentation: (ポスター発表)
• [Presentation] Folding of cysteine-rich peptides on phospholipid membranes. (2013)
  • Author(s): Hara1, T.; Purwati, E.M.; Wada, A.; Hasegawa, M.; Murata, M.; Aimoto, S.
  • Organizer: 4th Asia-Pacific International Peptide Symposium & 50th Japanese Peptide Symposium
  • Place of Presentation: 吹田市
• [Presentation] Purification of Transmembrane Peptides using RP-HPLC. (2013)
  • Author(s): Huang, Y.; Hara, T.; Murata M.
  • Organizer: 4th Asia-Pacific International Peptide Symposium & 50th Japanese Peptide Symposium
  • Place of Presentation: 吹田市
• [Presentation] ヒト心筋型脂肪酸結合タンパク質FABP3と蛍光プローブ1-anilinonaphtalene-8-sulphonic acidの複合体結晶構造解析 (2013)
  • Author(s): 廣瀬未果、松岡茂・石田英子・松岳大輔・原利明・溝端栄一・村上聡・井上豪・村田道雄・杉山成、
  • Organizer: 日本結晶学会 平成25年度年会
  • Place of Presentation: 熊本市
• [Presentation] Novel fatty acid analogues for the understanding of lipid-protein interactions. (2013)
  • Author(s): Hanako Ishida, Shigeru Sugiyama, Mika Hirose, Sebastien Lethu, Hikaru Ano, Fuminori Sato, Daisuke Matsuoka, Toshiaki Hara, Eiichi Mizohata, Tsuyoshi Inoue, Shigeru Matsuoka, Michio Murata.
  • Organizer: he International Conference on Structural Genomics 2013 (ICSG2013)
  • Place of Presentation: 札幌
• [Presentation] システインリッチペプチドの脂質膜上での酸化的フォールディング反応 (2013)
  • Author(s): 原利明、Euis Maras Purwati 、和田昭裕、長谷川慎、村田道雄、相本三郎
  • Organizer: 第13回 日本蛋白質科学会年会
  • Place of Presentation: 鳥取
• [Presentation] システインリッチペプチドの脂質膜上での酸化的フォールディング反応 (2012)
  • Author(s): 原利明、エイスマラスプルワティ、和田昭裕、長谷川慎、村田道雄、相本三郎
  • Organizer: 第13回日本蛋白質科学会年会
  • Place of Presentation: 鳥取(ポスター発表)
• [Presentation] 脂質膜を用いるシステインリッチペプチドのフォールディング反応 (2012)
  • Author(s): 原利明、Euis Maras Purwati 、和田昭裕、長谷川慎、村田道雄、相本三郎
  • Organizer: 第７回 日本ケミカルバイオロジー学会年回
  • Place of Presentation: 京都
• [Book] 原利明「化学合成ペプチド・タンパク質の調製と結晶構造解析」、杉山成編『タンパク質結晶の最前線』第II編 (2013)
  • Author(s): 原利明
  • Publisher: シーエムシー出版
• [Book] タンパク質結晶の最前線 第 II 編 (2013)
  • Author(s): 原利明 (杉山成 編)
  • Total Pages: 7
  • Publisher: シーエムシー出版
• [Remarks] ERATO村田脂質活性構造プロジェクト
  • URL: http://www.jst.go.jp/erato/murata/
• [Remarks] 蛋白質有機化学研究室 研究内容
  • URL: http://www.protein.osaka-u.ac.jp/organic/index.html