Study on the Glu-specific protease superfamily mediating dissociation of cell-cell adhesion

• Project/Area Number: 22592070
• Research Category: Grant-in-Aid for Scientific Research (C)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Functional basic dentistry
• Research Institution: Nagasaki University
• Principal Investigator: NEMOTO Yuko 長崎大学, 医歯薬学総合研究科, 准教授 (10164667)
• Co-Investigator(Kenkyū-buntansha): NEMOTO Takayuki 長崎大学, 大学院・医歯薬学総合研究科, 教授 (90164665) ; KIMURA Shigenobu 岩手医科大学, 歯学部, 教授 (10177917)
• Co-Investigator(Renkei-kenkyūsha): BABA Tomomi 長崎大学, 大学院・医歯薬学総合研究科, 助教 (60189727) ; KOBAYAKAWA Takeshi 長崎大学, 大学院・医歯薬学総合研究科, 教務職員 (10153587) ; ONO Toshio 長崎大学, 大学院・医歯薬学総合研究科, 研究員 (80050607) ; SHIMOYAMA Yu 岩手医科大学, 歯学部, 助教 (90453331)
• Project Period (FY): 2010 – 2012
• Project Status: Completed (Fiscal Year 2012)
• Budget Amount: ¥4,420,000 (Direct Cost: ¥3,400,000、Indirect Cost: ¥1,020,000); Fiscal Year 2012: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2011: ¥1,040,000 (Direct Cost: ¥800,000、Indirect Cost: ¥240,000); Fiscal Year 2010: ¥2,340,000 (Direct Cost: ¥1,800,000、Indirect Cost: ¥540,000)
• Keywords: セリンプロテアーゼ / プロテアーゼ / リコンビナントタンパク質 / 病原因子 / 黄色ブドウ球菌 / 活性化 / ジペプチジルペプチダーゼ / ブドウ球菌 / 酵素

Research Abstract

Shortening of the propeptide of Glu-specific serine peptidase(GluV8) family commonly occurred in an auto-catalytic manner. Accompanied with propeptide-shortening, the pro-mature junction became more susceptible toward the maturation enzymes. We demonstrated the in vitro Glu-specific endopeptidase activity of exfoliative toxin (ET) A for the first time, which was measured with Escherichia coli-expressed recombinant ETAs and synthetic peptidyl substrates. The animal model revealed that the proteolytic activity was indispensable for exfoliation of the skin. In addition, we determined characteristics of DPP11 and DPP7, and proposed a novel phylogenetic system of these family molecules.

Research Products

• [Journal Article] Discrimination based on Gly and Arg/Ser at Position 673 between Dipeptidyl-peptidase (DPP) 7 and DPP11, Widely Distributed DPPs in Pathogenic and Environmental Gram-Negative Bacteria (2013)
  • Author(s): Rouf, S.M.A., Ohara-Nemoto, Y., Hoshino, T., Fujiwara, T., Ono, T., Nemoto, T.K.
  • Journal Title: Biochimie Volume: 95 Issue: 4 Pages: 824-831
  • DOI: 10.1016/j.biochi.2012.11.019
  • NAID: 120006985514
  • Peer Reviewed
• [Journal Article] In vitro measurement of protease activity of exfoliative toxin A using synthetic peptide substrates (2013)
  • Author(s): Tachi, M.T., Ohara-Nemoto, Y., Baba, T.T., Kobayakawa, T., Fujita, S., Ikeda, T., Ayuse, T., Oi, K., Nemoto, T.K.
  • Journal Title: Acta Medica Nagasakiensia Volume: in press
  • Peer Reviewed
• [Journal Article] Rapid identification of Abiotrophia/Granulicatella species using by 16S rRNA based PCR and RFLP (2012)
  • Author(s): Shimoyama Yu
  • Journal Title: Interface Oral Health Science Pages: 206-208
  • Peer Reviewed
• [Journal Article] Maternal transmission of mutans and other oral streptococcal species (2012)
  • Author(s): Otake-Asakawa Asami
  • Journal Title: Interface Oral Health Science Pages: 198-200
  • Peer Reviewed
• [Journal Article] Asp- and Glu-specific novel dipeptidyl peptidase 11 of Porphyromonas gingivalis that ensures utilization of proteinaceous energy sources (2011)
  • Author(s): Ohara-Nemoto Y, Shimoyama Y, Kimura S, Kon A, Haraga H, Nemoto TK
  • Journal Title: J Biol Chem Volume: 286 Issue: 44 Pages: 38115-38127
  • DOI: 10.1074/jbc.m111.278572
  • Peer Reviewed
• [Journal Article] Amino acid residues modulating the activities of staphylococcal glutamyl endopeptidases (2010)
  • Author(s): Ono, T., Ohara-Nemoto, Y., Shimoyama, Y., Okawara, H., Kobayakawa, T., Baba, T., Kimura, S., Nemoto, T.K.
  • Journal Title: Biol Chem Volume: 391 Issue: 10 Pages: 1221-1232
  • DOI: 10.1515/bc.2010.116
  • NAID: 120006984895
  • Peer Reviewed
• [Journal Article] Amino acid residues modulating the activities of staphylococcal glutamyl endopeptidases. (2010)
  • Author(s): Ono Toshio
  • Journal Title: Biological Chemistry Volume: 391 Pages: 1221-1262
  • NAID: 120006984895
  • Peer Reviewed
• [Journal Article] Relationship between oral status and prevalence of periodontopathic bacteria on tongue of 85-years elderly. (2010)
  • Author(s): Kishi Mitsuo
  • Journal Title: Journal of Medical Microbiology Volume: 59 Pages: 1354-1359
  • Peer Reviewed
• [Journal Article] A novel aspartate-specific dipeptidylpeptidase produced from Porphyromonas endodontalis. (2010)
  • Author(s): Kimura Sigenobu
  • Journal Title: Interface Oral Health Science Volume: 2009 Pages: 265-266
  • Peer Reviewed
• [Journal Article] In vitro measurement of protease activity of exfoliative toxin A using synthetic peptide substrates.
  • Author(s): Tachi, M.T., Ohara-Nemoto, Y., Baba, T.T., Kobayakawa, T., Fujita, S., Ikeda, T., Ayuse, T., Oi, K., Nemoto, T.K.
  • Journal Title: Acta Medica Nagasakiensia Volume: (in press)
  • Peer Reviewed
• [Presentation] Dipeptide production and assimilation in Porphyromonas gingivalis by four dipeptidyl peptidases. Nagasaki University Global COE symposium (2012)
  • Author(s): Rouf, S.M.A., Ohara-Nemoto, Y., Nemoto, T.K.
  • Organizer: The 6th Nagasaki Symposium on Tropical and Emerging Infectious Diseases and The 11th Nagasaki-Singapore Medical Symposium
  • Place of Presentation: Nagasaki, Japan
  • Year and Date: 2012-12-10
• [Presentation] Expression and Characterization of Dipeptidyl- Peptidases of Porphyromonas gingivalis (2012)
  • Author(s): Rouf, S.M.A., Ohara-Nemoto, Y, Yanase, A., Ono, T., Nemoto, T.K
  • Organizer: 1st International Conference on Porphyromonas gingivalis and Related Bacterial Species
  • Place of Presentation: Nagasaki, Japan
  • Year and Date: 2012-08-27
• [Presentation] Classification and essential amino acid residues of the dipeptidyl peptidase 7/11 family in the Cytophaga- Flavobacterium-Bacteroides group (2012)
  • Author(s): Ohara-Nemoto, Y., Rouf, S.M.A., Hoshino, T., Fujiwara, T., Shimoyama, Y., Kimura, S., Ono, T., Nemoto, T.K.
  • Organizer: 112th General Meeting of ASM
  • Place of Presentation: San Francisco, USA
  • Year and Date: 2012-06-18
• [Presentation] 表皮剥脱毒素 ETA の生物活性とプロテアーゼ活性の相関性 (2012)
  • Author(s): 達聖月,小早川健,根本孝幸,馬場友巳,根本優子
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎市
  • Year and Date: 2012-03-29
• [Presentation] 表皮剥脱毒素ETAの生物活性とプロテアーゼ活性の相関性 (2012)
  • Author(s): 達聖月
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎ブリックホール(長崎市)
  • Year and Date: 2012-03-29
• [Presentation] Porphyromonas属菌が産生するAsp/Glu特異的新規ジペプチジルペプチダーゼ(DPP)11の同定と基質特異性の検討 (2012)
  • Author(s): 根本孝幸, SMAラオフ, 小野俊雄,下山佑,木村重信,根本優子
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎市
  • Year and Date: 2012-03-27
• [Presentation] Porphyromonas属菌が産生するAsp/Glu特異的新規ジペプチジルペプチダーゼ(DPP)11の同定と基質特異性の検討 (2012)
  • Author(s): 根本孝幸
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎ブリックホール(長崎市)
  • Year and Date: 2012-03-27
• [Presentation] DPP7と新規Asp/Glu特異的DPP11を含むS46ペプチダーゼファミリーの再分類 (2012)
  • Author(s): Rouf W.A.Shakh
  • Organizer: 第85回日本細菌学会総会
  • Place of Presentation: 長崎ブリックホール(長崎市)
  • Year and Date: 2012-03-27
• [Presentation] Dipeptide production and assimilation in Porphyromonas gingivalis by four dipeptidyl peptidases (2012)
  • Author(s): Rouf, Shakh M. A., Ohara-Nemoto, Y., and Nemoto, T. K.
  • Organizer: Nagasaki University Global COE symposium
  • Place of Presentation: Nagasaki, Japan
• [Presentation] Expression and Characterization of Dipeptidyl-Peptidases of Porphyromonas gingivalis (2012)
  • Author(s): Rouf, Shakh M. A., Ohara-Nemoto, Y., Yanase, A., Ono T., and Nemoto, T. K.
  • Organizer: 1st International Conference on Porphyromonas gingivalis and Related Bacterial Species
  • Place of Presentation: Nagasaki, Japan
• [Presentation] Classification and essential amino acid residues of the dipeptidyl peptidase 7/11 family in the Cytophaga-Flavobacterium-Bacteroides group (2012)
  • Author(s): Ohara-Nemoto, Y., Abdur-Rouf, S. M., Hoshino, T., Fujiwara, T., Shimoyama, Y., Kimura, S., Ono, T., and Nemoto, T. K.
  • Organizer: 112th General Meeting of American Society for Microbiology
  • Place of Presentation: San Francisco, CA, USA
• [Presentation] Colonization of Porphyromonas gingivalis in the plaques from mothers and their children (2012)
  • Author(s): Kimura, M., Shimoyama, Y., Ohara-Nemoto, Y., Kimura, S. and Tanaka, M.
  • Organizer: 11th Congress of the European Academy of Paediatric Dentistry
  • Place of Presentation: Strasbourg, France
• [Presentation] Classification of the S46 (DPP7/DPP11) family members and identification of a subtype of DPP11 (2012)
  • Author(s): 根本孝幸，ラオフ アブドユー，小野俊夫，下山佑，木村重信，根本優子
  • Organizer: 第54回歯科基礎医学学術大会
  • Place of Presentation: 郡山市
• [Presentation] Substrate specificity of four dipeptidyl peptidases from Porphyromonas gingivalis and the dipeptide repertoire produced by the bacterium (2012)
  • Author(s): 柳瀬絵美，ラオフ アブドユー，小野俊夫，根本優子，根本孝幸
  • Organizer: 第54回歯科基礎医学学術大会
  • Place of Presentation: 郡山市
• [Presentation] Porphyromonas属菌が産生するAsp/Glu特異的新規ジペプチジルペプチダーゼ(DPP)11 (2011)
  • Author(s): 根本優子,下山佑,木村重信,根本孝幸
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 岐阜市
  • Year and Date: 2011-10-01
• [Presentation] Porphyromonas gingivalisDPP11の酵素活性とAsp/Glu特異性を規定するArg670 (2011)
  • Author(s): 根本孝幸,木村重信,下山佑,木村重信,根本優子
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 岐阜市
  • Year and Date: 2011-10-01
• [Presentation] 表皮剥奪毒素ETA N 末端領域の生物活性に及ぼす作用 (2011)
  • Author(s): 達聖月,根本優子,馬場友巳,小早川健,藤田修一,池田通,大井久美子,根本孝幸
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 岐阜市
  • Year and Date: 2011-10-01
• [Presentation] Porphyromonas gingivalis DPP11の酵素活性とAsp/Glu特異性を規定するArg670 (2011)
  • Author(s): 根本孝幸
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 国立京都国際会館(京都市)
  • Year and Date: 2011-10-01
• [Presentation] Porphyromonas属菌が産生するAsp/Glu特異的新規ジペプチジルペプチダーゼ(DPP)11 (2011)
  • Author(s): 根本優子
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 長良川国際会議場(岐阜市)
  • Year and Date: 2011-10-01
• [Presentation] 表皮剥奪毒素ETA N末端領域の生物活性に及ぼす作用 (2011)
  • Author(s): 達聖月
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 長良川国際会議場(岐阜市)
  • Year and Date: 2011-10-01
• [Presentation] 骨芽細胞に発現するアクアポリン (2011)
  • Author(s): 馬場友巳
  • Organizer: 第53回歯科基礎医学学術大会
  • Place of Presentation: 長良川国際会議場(岐阜市)
  • Year and Date: 2011-10-01
• [Presentation] Biochemical characterization of DPP11 : Importance of arginine670 for the enzymatic activity specific for the P1-position acidic residue (2011)
  • Author(s): 根本孝幸
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館(京都市)
  • Year and Date: 2011-09-24
• [Presentation] A novel Asp and Glu specific dipeptitidyl peptidase from Porphyromonas endodontalis (2011)
  • Author(s): Tetuo Fumi
  • Organizer: 38th Asia Pacific Dental Students Association Congress
  • Place of Presentation: Rama Garden Hotel Bangkok (Bangkok, Thailand)
  • Year and Date: 2011-08-17
• [Presentation] Auto-catalytic propeptide processing facilitates the final hetero-catalytic maturation of glutamyl endopeptidase GluV8 (2011)
  • Author(s): Ohara-Nemoto, Y, Rouf, S.M.A, Shimoyama, Y., Kimura, S., Ono, T., Nemoto, T.K.
  • Organizer: 111th General Meeting of AmericanSociety for Microbiology
  • Place of Presentation: New Orleans, USA
  • Year and Date: 2011-05-24
• [Presentation] Auto-catalytic propeptide processing facilitates the final hetero-catalytic maturation of glutamyl endopeptidase GluV8 (2011)
  • Author(s): Ohara-Nemoto Yuko
  • Organizer: 111th General Meeting of American Society for Microbiology
  • Place of Presentation: The Ernest N.Morial Convention Center (New Orleans, USA)
  • Year and Date: 2011-05-24
• [Presentation] Auto-catalytic processing of prosequence prior to maturation of Staphylococcal glutamyl endopeptidases. (2010)
  • Author(s): Shakh M.A.Rouf
  • Organizer: 第33回日本分子生物学会年会,第83回日本生化学会大会合同大会
  • Place of Presentation: 神戸市
  • Year and Date: 2010-12-10
• [Presentation] 新規V8プロテアーゼファミリー遺伝子のクローニングとV8プロテアーゼ活性に重要なN末バリンと185-195領域の同定 (2010)
  • Author(s): 根本孝幸
  • Organizer: 第52回歯科基礎医学学術大会
  • Place of Presentation: 東京都
  • Year and Date: 2010-09-21
• [Presentation] Structural chemistry of Staphylococcal aureus glutamyl endopeptidases. (2010)
  • Author(s): Ohara-Nemoto Yuko
  • Organizer: 88th General Session of International Association for Dental Research
  • Place of Presentation: Barcelona, Spain
  • Year and Date: 2010-09-02
• [Book] NVS and staphylococci in the oral cavity, a cause of infective endocarditis (2012)
  • Author(s): Ohara-Nemoto, Y., Kimura, S., Nemoto, T.K.
  • Publisher: In Endocarditis InTech
• [Book] Pathogenic factors of P. Gingivalis and the host defence mechanismsn Pathogenesis and Treatment of Periodontitis. (2012)
  • Author(s): Kimura, S., Ohara-Nemoto, Y., Shimoyama, Y., Ishikawa, T., Sasaki, M.
  • Publisher: InTech
• [Book] Endocarditis (2012)
  • Author(s): Ohara-Nemoto, Y., Kimura, S., Nemoto, T.K.
  • Total Pages: 22
  • Publisher: NVS and staphylococci in the oral cavity, a cause of infective endocarditis
• [Book] Pathogenesis and Treatment of Periodontitis (2012)
  • Author(s): Kimura, S., Ohara-Nemoto, Y., Shimoyama, Y., Ishikawa, T., Sasaki, M.
  • Total Pages: 16
  • Publisher: Pathogenic factors of P. gingivalis and the host defence mechanisms
• [Book] Endocarditis (2012)
  • Author(s): Ohara-Nemoto Yuko
  • Publisher: INTECK
• [Book] Pathogenesis and Treatment of Periodontitis (2012)
  • Author(s): Kimura Shigenobu
  • Publisher: INTECK
• [Remarks]
  • URL: http://www.de.nagasaki-u.ac.jp/education/dept_omb.html
• [Remarks] 長崎大学歯学部口腔分子生化学
• [Remarks]
  • URL: http://naosite.lb.nagasaki-u.ac.jp/dspace/