Translational research on a drug delivery system using substrate binding domain derived from bacterial collagenases

• Project/Area Number: 20590452
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Bacteriology (including Mycology)
• Research Institution: Kitasato University
• Principal Investigator: MATSUSHITA Osamu Kitasato University, 医学部, 教授 (00209537)
• Co-Investigator(Kenkyū-buntansha): KOIDE Takaki 早稲田大学, 先進理工学部, 教授 (70322253) ; TAMAI Eiji 松山大学, 薬学部, 准教授 (40333512) ; MIYATA Shigeru 香川大学, 医学部, 講師 (90314913)
• Co-Investigator(Renkei-kenkyūsha): MIMAMI Junzaburo 香川県立保健医療大学, 保健医療学部, 教授 (40157566)
• Project Period (FY): 2008 – 2010
• Project Status: Completed (Fiscal Year 2010)
• Budget Amount: ¥4,810,000 (Direct Cost: ¥3,700,000、Indirect Cost: ¥1,110,000); Fiscal Year 2010: ¥130,000 (Direct Cost: ¥100,000、Indirect Cost: ¥30,000); Fiscal Year 2009: ¥1,430,000 (Direct Cost: ¥1,100,000、Indirect Cost: ¥330,000); Fiscal Year 2008: ¥3,250,000 (Direct Cost: ¥2,500,000、Indirect Cost: ¥750,000)
• Keywords: ガス壊疽菌群 / 毒素 / 構造活性相関 / コラーゲン結合ドメイン / 成長因子 / 骨新生 / 鼓膜閉鎖術 / 人工皮膚 / コラゲナーゼ / 上皮細胞成長因子 / 鼓膜穿孔 / 血管内皮細胞成長因子 / 血管新生 / 再生医療 / スピン標識ペプチド / 表皮成長因子 / 重層扁平上皮

Research Abstract

'Flesh-eating bacteria' produce collagenases to hydrolyze collagen fibrils in tissue. These enzymes possess domain(s) to bind to this insoluble substrate (CBD). We identified amino acid residues responsible for the binding. We also showed that CBD binds preferentially to the under-wound region in the collagen molecule, and that the binding is unidirectional. Ca^<2+> binds to the linker region of CBD. We showed that the binding modifies CBD into a more stable and more efficient conformation. Finally, we showed the potential benefits of matrix anchoring of various bioactivity substances using CBD, which are applicable to various therapies in regenerative medicine.

Research Products

• [Journal Article] Monthly administration of a novel PTH-vollagen binding domain fusion protein is anabolic in mice. Calcif. (2011)
  • Author(s): Ponnapakkam T., Katikaneni, R., Miller, E., Ponnapakkam, A., Suda, H., Miyata, S., Suva, L.J., JSakon, J., Matsushita, O., Gensure, R.C.
  • Journal Title: Tissue Int. in press
  • Peer Reviewed
• [Journal Article] Development of a high-throughput screening system for the compounds that inhibit collagen-protein interactions. (2009)
  • Author(s): Okano-Kosugi, H., Matsushita, O., Asada, S., Herr, A.B., Kitagawa, K., Koide, T.
  • Journal Title: Anal.Biochem. 394 Pages: 125-131
  • Peer Reviewed
• [Journal Article] Ca2^+-induced linker transformation leads to a compact and rigid collagen-binding domain of Clostridium histolyticum collagenase. (2009)
  • Author(s): Philominathan, S.T., Matsushita, O., Gensure, R., Sakon, J.
  • Journal Title: FEBS J. 276 Pages: 3589-3601
  • Peer Reviewed
• [Journal Article] Unidirectional binding of clostridial collagenase to triple helical substrates. (2009)
  • Author(s): Philominathan, S.T., Koide, T., Hamada, K., Yasui, H., Seifert, S., Matsushita, O., Sakon, J.
  • Journal Title: J.Biol.Chem. 284 Pages: 10868-10876
  • Peer Reviewed
• [Journal Article] Unidirectional binding of clostridial collagenase to triple helical substrates. (2009)
  • Author(s): Philominathan ST, Koide T, Hamada K, Yasui H, Seifert S, Matsushita O, Sakon J.
  • Journal Title: J.Biol.Chem. 284 Pages: 10868-10876
  • Peer Reviewed
• [Journal Article] Ca^<2+>-induced linker transformation leads to a compact and rigid collagen-binding domain of Clostridium histolyticum collagenase. (2009)
  • Author(s): Philominathan ST, Matsushita O, Gensure R, Sakon J.
  • Journal Title: FEBS J. 276 Pages: 3589-3601
  • Peer Reviewed
• [Journal Article] Development of a high-throughput screening system for the compounds that inhibit collagen-protein interactions. (2009)
  • Author(s): Okano-Kosugi H, Matsushita O, Asada S, Herr AB, Kitagawa K, Koide T.
  • Journal Title: Anal.Biochem. 394 Pages: 125-131
  • Peer Reviewed
• [Journal Article] 1H, 13C and 15N resonace assignments of Ca2^+-bound collagen-binding domain derived from a clostridial collagenase. (2008)
  • Author(s): Philominathan, S.T.L., Matsushita, O., Jordan, J.B., Sakon, J.
  • Journal Title: Biomol.NMR Assign. 2 Pages: 127-129
  • Peer Reviewed
• [Journal Article] High-level expression of his-tagged clostridial collagenase in Clostridium perfringens. (2008)
  • Author(s): Tamai, E., Miyata, S., Tanaka, H., Nariya, H., Suzuki,M., Matsushita, O., Hatano, N., Okabe, A.
  • Journal Title: Appl.Microbiol.Biotechnol. 80 Pages: 627-635
  • Peer Reviewed
• [Journal Article] High-level expression of his-tagged clostridial collagenase in Clostridium perfringens. (2008)
  • Author(s): Tamai, E., Miyata, S., Tanaka, H., Nariya, H., Suzuki, M., Matsushita, O., Hatano, N., Okabe, A.
  • Journal Title: Appl. Microbiol. Biotechnol. 80 Pages: 627-635
  • Peer Reviewed
• [Journal Article] ^1H,^<13>C and ^<15>N resonace assignments of Ca^<2+> bound collagen-binding domain derived from a clostridial collagenase (2008)
  • Author(s): Philominathan, S. T. L., Matsushita, O., Jordan, J. B., Sakon, J.
  • Journal Title: Biomol. NMR Assign. 2 Pages: 127-129
  • Peer Reviewed
• [Presentation] From bacterial toxin to drug. (2010)
  • Author(s): Matsushita O., Collagen anchoring
  • Organizer: 11th Joint Symposium Robert Koch Institute and The Kitasato Institute Kitasato University
  • Place of Presentation: 東京都港区
  • Year and Date: 2010-10-26
• [Presentation] Collagen anchoring : From bacterial toxin to drug (2010)
  • Author(s): Osamu Matsushita
  • Organizer: 11th Joint Symposium Robert Koch Institute and The Kitasato Institute Kitasato University
  • Place of Presentation: 北里大学薬学部コンベンションホール(東京都)
  • Year and Date: 2010-10-26
• [Presentation] ガス壊疽菌群コラゲナーゼの基質結合ドメインと再生医療への応用 (2010)
  • Author(s): 松下治、西望、小杉日登美、小出隆規、安達栄治郎、坂本恵子、服部雅一
  • Organizer: 第57回トキシンシンポジウム
  • Place of Presentation: 滋賀県長浜市
  • Year and Date: 2010-07-16
• [Presentation] ガス壊疽菌群コラゲナーゼの基質結合ドメインと再生医療への応用 (2010)
  • Author(s): 松下治、西望、小杉日登美、小出隆規、安達栄治郎、坂本恵子、服部雅一
  • Organizer: 第57回トキシンシンポジウム
  • Place of Presentation: 長浜ロイヤルホテル(滋賀県)
  • Year and Date: 2010-07-16
• [Presentation] コラーゲン結合ドメインの構造解析と薬物送達システムへの応用 (2010)
  • Author(s): 松下治、宮田茂
  • Organizer: 第83回日本細菌学会
  • Place of Presentation: 神奈川県横浜市
  • Year and Date: 2010-03-27
• [Presentation] ガス壊疽菌群コラゲナーゼの基質結合様式と積層型人工組織作製への応用 (2009)
  • Author(s): 松下治
  • Organizer: 第56回トキキンシンポジウム
  • Place of Presentation: 岐阜県岐阜市
  • Year and Date: 2009-08-27
• [Presentation] Substrate recognition of collagen-binding domains derived from bacterial collagenases. (2009)
  • Author(s): Matsushita O., Nishi N., Koide T., Philominathan S.T.L., Sakon J., Gensure R.C., Iwashiro H., Adachi E.
  • Organizer: 8th Pan-pacific connective tissue societies symposium, 41st Annual meeting of the Japanese society of connective tissue research, 56th Annual meeting of the Japan matrix club
  • Place of Presentation: 神奈川県逗子市
  • Year and Date: 2009-06-04
• [Presentation] Substrate recognition of collagen-binding domains derived from bacterial collagenases (2009)
  • Author(s): Matsushita O, Nishi N, Koide T, Philominathan STL, Sakon J Gensure RC, Iwashiro H, Adachi E
  • Organizer: 8th Pan-pacific connective tissue societies symposium, 41st Annual meeting of the Japanese society of connective tissue research, 56th Annual meeting of the Japan matrix club
  • Place of Presentation: 神奈川県逗子市
  • Year and Date: 2009-06-04
• [Presentation] ガス壊疽菌群コラゲナーゼの基質認識機構と再生医学への応用 (2009)
  • Author(s): 松下治、小出隆規、安達栄治郎
  • Organizer: 第82回日本細菌学会総会
  • Place of Presentation: 愛知県名古屋市
  • Year and Date: 2009-03-12
• [Presentation] ガス壊疽菌群コラゲナーゼのセグメント2の構造解析 (2008)
  • Author(s): 松下治
  • Organizer: 第91回日本細菌学会関東支部総会
  • Place of Presentation: 千葉県長生郡長塚町
  • Year and Date: 2008-10-23
• [Remarks] ホームページ等
  • URL: http://web.med.kitasato-u.ac.jp/edures/microbio/toxin.html
• [Remarks]
  • URL: http://web.med.kitasato-u.ac.jp/edures/microbio/toxin.html
• [Patent(Industrial Property Rights)] 粘膜、上皮および鼓膜の少なくとも一つの再生材および再生評価方法 (2010)
  • Inventor(s): 宮下武憲、森望、西望、安達栄治郎、松下治、服部雅一、伊藤周平、坂本恵子
  • Industrial Property Rights Holder: 国立大学法人香川大学、学校法人北里研究所
  • Industrial Property Number: 2010-110694
  • Filing Date: 2010-05-12
• [Patent(Industrial Property Rights)] 積層型高密度培養人工組織の製造方法及び積層型高密度培養人工組織 (2010)
  • Inventor(s): 安達栄治郎、松下治、岩城啓修、細谷智、西望
  • Industrial Property Rights Holder: 学校法人北里研究所、国立大学法人香川大学
  • Filing Date: 2010-01-28
  • Overseas
• [Patent(Industrial Property Rights)] 粘膜、上皮および鼓膜の少なくとも-つの再生材および再生評価方法 (2010)
  • Inventor(s): 宮下武憲、森望、西望、安達栄治郎、松下治、服部雅一、伊藤周平、坂本恵子
  • Industrial Property Rights Holder: 香川大学 北里大学
  • Industrial Property Number: 2010-110694
  • Filing Date: 2010-05-12
• [Patent(Industrial Property Rights)] 積層型高密度培養人工組織の製造方法及び積層型高密度培養人工組織 (2010)
  • Inventor(s): 安達栄治郎、松下治、岩城啓修、細谷智、西望
  • Industrial Property Rights Holder: 学校法人北里研究所・国立大学法人香川大学
  • Filing Date: 2010-01-28
  • Overseas
• [Patent(Industrial Property Rights)] 積層型高密度培養人工組織の製造方法及び積層型高密度培養人工組織 (2009)
  • Inventor(s): 安達栄治郎、松下治、岩城啓修、細谷智、西望
  • Industrial Property Rights Holder: 学校法人北里研究所、国立大学法人香川大学
  • Industrial Property Number: 2009-017475
  • Filing Date: 2009-01-19
• [Patent(Industrial Property Rights)] 積層型高密度培養人工組織の製造方法及び積層型高密度培養人工組織 (2009)
  • Inventor(s): 安達栄治郎, 松下治, 岩城啓修, 細谷 智, 西 望
  • Industrial Property Rights Holder: 学校法人北里研究所, 国立大学法人香川大学
  • Industrial Property Number: 2009-017475
  • Filing Date: 2009-01-19