Study on the Structures, Functions and Applications of Sphingolipid-metabolizing machinery

• Project/Area Number: 21380066
• Research Category: Grant-in-Aid for Scientific Research (B)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Applied biochemistry
• Research Institution: Kyushu University
• Principal Investigator: ITO Makoto 九州大学, 大学院・農学研究院, 教授 (40253512)
• Co-Investigator(Kenkyū-buntansha): OKINO Nozomu 九州大学, 大学院・農学研究院, 准教授 (90363324) ; KAKUTA Yoshimitsu 九州大学, 大学院・農学研究院, 准教授 (00314360)
• Project Period (FY): 2009 – 2011
• Project Status: Completed (Fiscal Year 2011)
• Budget Amount: ¥18,590,000 (Direct Cost: ¥14,300,000、Indirect Cost: ¥4,290,000); Fiscal Year 2011: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2010: ¥5,330,000 (Direct Cost: ¥4,100,000、Indirect Cost: ¥1,230,000); Fiscal Year 2009: ¥7,930,000 (Direct Cost: ¥6,100,000、Indirect Cost: ¥1,830,000)
• Keywords: 糖鎖生物学 / 酵素利用学 / スフィンゴ(糖)脂質 / 代謝 / 結晶構造 / 糖脂質 / 酵素 / 生体分子 / 糖鎖 / 菌類 / エンドグリコセラミダーゼ / EGCrP / 感染症 / スフィンゴ脂質 / 代謝酵素 / X線結晶構造解析 / 生物機能 / セラミダーゼ

Research Abstract

In this study, we explored the structures and functions of novel sphingolipid-metabolizing enzymes(ceramidase, sphingolipid ceramide N-deacylase, Hex1, NgaP, EGCrP1, EGALC), which had been identified and cloned in our laboratory. Interestingly, both ceramidase and sphingolipid ceramide N-deacylase are a zinc-dependent amidohydrolase. The cleavage of N-acyl linkage of ceramide by ceramidase follows a similar mechanism as observed for the zinc-dependent carboxypeptidases. Hex1 and NgaP were found to cleave the terminal

Research Products

• [Journal Article] Quality control of fungus-specific glucosylceramide in Cryptococcus neoformans by endoglycoceramidase-related protein 1(EGCrP1) (2012)
  • Author(s): Ishibashi Y, Okino N(6人中4番目), Ito M(6人中6番目)
  • Journal Title: J. Biol. Chem Volume: 287 Pages: 368-381
  • Peer Reviewed
• [Journal Article] Quality control of fungus-specific glucosylceraraide in Cryptococcus neoformans by endoglycoceramidase-related protein 1 (EGCrP1) (2012)
  • Author(s): Y.Ishibashi, et al
  • Journal Title: Journal of Biological Chemistry Volume: 287 Issue: 1 Pages: 368-381
  • DOI: 10.1074/jbc.m111.311340
  • Peer Reviewed
• [Journal Article] Qualitative and quantitative cellular glycomics of glycosphingolipids based on rhodococcal endoglycoceramidase-assisted glycan cleavage, glycoblotting-assisted sample preparation, and matrix-assisted laser desorption ionization tandem time-of-flight mass (2011)
  • Author(s): Fujitani N, Ito M(9人中8番目)
  • Journal Title: J. Biol. Chem Volume: 286 Pages: 41669-41679
  • Peer Reviewed
• [Journal Article] Molecular cloning and catalytic mechanism of a novel glycosphingolipid-degradingβ-N-acetylgalactosaminidase from Paenibacillus sp (2011)
  • Author(s): Sumida T, Fujimoto K, Ito M
  • Journal Title: TS12. J. Biol. Chem Volume: 286 Pages: 14065-14072
  • Peer Reviewed
• [Journal Article] Qualitative and quantitative cellular glycomics of glycosphingolipids based on Rhodococcal endoglycoceramidase-assisted glycan cleavage, glycoblotting-assisted sample preparation, and matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry analysis (2011)
  • Author(s): N.Fujitani, et al
  • Journal Title: Journal of Biological Chemistry Volume: 286 Pages: 41669-41769
  • Peer Reviewed
• [Journal Article] Molecular cloning and catalytic mechanism of a novel glycosphingolipid-degrading β-N-acetylgalactosminidase from Paenibacillus sp. TS12 (2011)
  • Author(s): T.Sumida, et al
  • Journal Title: Journal of Biological Chemistry Volume: 286 Pages: 14065-14072
  • Peer Reviewed
• [Journal Article] Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation (2010)
  • Author(s): Togayachi A, Ito M(15人中11番)
  • Journal Title: PNAS Volume: 107 Pages: 11900-11905
  • Peer Reviewed
• [Journal Article] Expression, purification, and characterization of a recombinant neutral ceramidase from Mycobacterium tuberculosis. (2010)
  • Author(s): Okino N, et al.
  • Journal Title: Biosci.Biotechnol.Biochem. Volume: 74(2) Pages: 316-321
  • NAID: 10027551401
  • Peer Reviewed
• [Journal Article] Lack of lacto/neolacto-glycolipids enhances the formation of glycolipid-enriched microdomains, facilitating B cell activation. (2010)
  • Author(s): Togayachi A, et al.
  • Journal Title: Proc.Natl.Acad.Sci.USA Volume: 107(26) Pages: 11900-11905
  • Peer Reviewed
• [Journal Article] β-Galactosyltransferase-5 is a lactosylceramide synthase essential for □mouse extra-embryonic development. (2010)
  • Author(s): Nishie T, et al.
  • Journal Title: Glycobiology Volume: 20(10) Pages: 1311-1322
  • Peer Reviewed
• [Journal Article] Simultaneous quantification of glucosylceramide and galactosylceramide by normal-phase HPLC using O-phatalaldehyde derivatives prepared with sphingolipid ceramide N-deacylase (2009)
  • Author(s): Zama K, Okino N(8人中7番), Ito M(8人中8番)
  • Journal Title: Glycobiology Volume: 19 Pages: 767-775
  • Peer Reviewed
• [Journal Article] Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endogalactosylceramidase (2009)
  • Author(s): Ishibashi Y, Okino N(9人中7番), Ito M(9人中9番)
  • Journal Title: Glycobiology Volume: 19 Pages: 797-807
  • Peer Reviewed
• [Journal Article] Zebrafish and mouseα2, 3-sialyltransferases responsible for synthesizing GM4 ganglioside (2009)
  • Author(s): Chisada S, Okino N(16人中14番目), Ito M.(16人中16番目)
  • Journal Title: J. Biol. Chem Volume: 284 Pages: 30534-30546
  • Peer Reviewed
• [Journal Article] Molecular cloning and crystal structural analysis of a novelβ-N-acetylhexosaminidase from Paenibacillus sp. TS12 capable of degrading glycosphingolipids (2009)
  • Author(s): Sumida T, Ishii R, Yanagisawa T, Yokoyama S, Ito M
  • Journal Title: J. Mol. Biol. Volume: 392 Pages: 87-99
  • Peer Reviewed
• [Journal Article] Mechanical insights into the hydrolysis and synthesis of ceramide by neutral ceramidase (2009)
  • Author(s): Inoue T, Okino N(12人中2番目First equal contribution), Kakuta Y(12人中3番目), Ito M(12人中12番目)
  • Journal Title: J. Biol. Chem Volume: 284 Pages: 9566-9577
  • Peer Reviewed
• [Journal Article] Mechanical insights into the hydrolysis and synthesis of ceramide by neutral ceramidase (2009)
  • Author(s): Inoue, et al.
  • Journal Title: Journal of Biological Chemistry 284 Pages: 9566-9577
  • Peer Reviewed
• [Journal Article] Tamavidins--novel avidin-like biotin-binding proteins from the Tamogitake mushroom (2009)
  • Author(s): Takakura, et al
  • Journal Title: FEBS J 276 Pages: 1383-1397
  • Peer Reviewed
• [Journal Article] Simultaneous quantification of glucosylceramide and galactosylceramide by normal-phase HPLC using O-phtalaldehyde derivatives prepared with sphingolipid ceramide N-deacylase (2009)
  • Author(s): Zama, et al.
  • Journal Title: Glycobiology 19 Pages: 767-775
  • Peer Reviewed
• [Journal Article] Transglycosylation-based fluorescent labeling of 6-gala series glycolipids by endoglycoceramidase (2009)
  • Author(s): Ishibashi, et al.
  • Journal Title: Glycobiology 19 Pages: 797-807
  • Peer Reviewed
• [Journal Article] Crystal structure of a/b-galacoside a2, 3-sialyltransferase from a luminous marine bacterium, Photobacterium phosphoreum (2009)
  • Author(s): Iwatani, et al.
  • Journal Title: FEBS Lett 583 Pages: 2083-2087
  • Peer Reviewed
• [Journal Article] Ca2+/calmodulin-dependent protein kinase phosphatase(CaMKP)is indispensable for normal embryogenesis in zebrafish, Danio rerio (2009)
  • Author(s): Sueyoshi, et al.
  • Journal Title: Arch Biochem Biophys 488 Pages: 48-59
  • Peer Reviewed
• [Journal Article] Characterization and application of carbohydrate-binding modules of b-1, 3-xylanase XUL4. (2009)
  • Author(s): Kiyohara, et al.
  • Journal Title: J Biochem 140 Pages: 369-373
  • Peer Reviewed
• [Journal Article] Molecular cloning and crystal structural analysis of a novel b-N-acetylhexosaminidase from Paenibacillus sp. TS12 capable of degrading glycosphingoliids (2009)
  • Author(s): Sumida, et al.
  • Journal Title: J Mol Biol 392 Pages: 87-99
  • Peer Reviewed
• [Journal Article] Zebrafish and mouse a2, 3-sialyltransferases responsible for synthesizing GM4 ganglioside (2009)
  • Author(s): Chisada, et al.
  • Journal Title: Journal of Biological Chemistry 284 Pages: 30534-30546
  • Peer Reviewed
• [Journal Article] A novel fucosyl glycosphingolipid of brine shrimp that is highly sensitive to endoglycoceramidase (2009)
  • Author(s): Xu, et al.
  • Journal Title: Glycobiology 141 Pages: 1-7
  • Peer Reviewed
• [Presentation] Endoglycoceramidase and its related protein EGCrP1 (2011)
  • Author(s): M.Ito
  • Organizer: 3rd Asian Communications of Glycobiology and Glycotechnoloey
  • Place of Presentation: Shanghai, China(招待講演)
  • Year and Date: 2011-10-29
• [Presentation] Biological significance of non-lysosomal degradation of glucosylceramide (2011)
  • Author(s): M.Ito
  • Organizer: Japan-Netherland Glycobiology Symposium
  • Place of Presentation: Nagoya, Japan(招待講演)
  • Year and Date: 2011-10-09
• [Presentation] Lipid metabolism in thraustochytrids (2011)
  • Author(s): M.Ito
  • Organizer: The 30th Naito Conference
  • Place of Presentation: Sapporo, Japan(招待講演)
  • Year and Date: 2011-07-01
• [Presentation] Endoglycoceramidase and its related protein EGCrP1 (2011)
  • Author(s): Ito M
  • Organizer: 3rd Asian Communications of Glycobiology and Glycotechnology
• [Presentation] Biological significance of non-lysososmal degradation of glucosylceramide (2011)
  • Author(s): Ito M
  • Organizer: 日蘭糖鎖科学シンポジウム
  • Place of Presentation: 名古屋大学、名古屋市
• [Presentation] グルコシルセラミドの品質管理と細胞分裂に関与する2つのEGCrPパラログ (2011)
  • Author(s): 伊東信、石橋洋平、伊藤友治、池田和貴、沖野望、合田初美、坂口圭史、田口良
  • Organizer: 第84回日本生化学会大会
  • Place of Presentation: 国立京都国際会館
• [Presentation] X-ray structure ofsphingolipid ceramide N-deacylase (2011)
  • Author(s): Okino N, Kakuta Y, Kato-Unoki Y, Sueyoshi N, Ito M
  • Organizer: The 31st Naito Conference
  • Place of Presentation: 北海道札幌市
• [Presentation] 全ての糖脂質から糖鎖を切り離す酵素の開発 (2010)
  • Author(s): 伊東信
  • Organizer: 第8回糖鎖科学コンソーシアムシンポジウム
  • Place of Presentation: 東京コンファレンスセンター、東京
  • Year and Date: 2010-11-30
• [Presentation] 全ての糖脂質から糖鎖を切り離す酵素の開発 (2010)
  • Author(s): 伊東信
  • Organizer: 糖鎖科学コンソーシアムシンポジウム
  • Place of Presentation: 東京
  • Year and Date: 2010-11-30
• [Presentation] The crystal structure of sphingolipid ceramide N-deacylase that catalyzes the cleavage and creation of an N-acyl linkage of various glycosphingolipids (2010)
  • Author(s): Okino N, et al.
  • Organizer: International Carbohydrate Symposium (ICS)
  • Place of Presentation: Makuhari, Chiba, Japan
  • Year and Date: 2010-08-05
• [Presentation] Discovery of a novel glucosylceramidase involved in the glucosylceramide metabolism in pathogenic fungi (2010)
  • Author(s): Ishibashi Y, et al.
  • Organizer: International Carbohydrate Symposium (ICS)
  • Place of Presentation: Makuhari, Chiba, Japan
  • Year and Date: 2010-08-05
• [Presentation] Pathogenic vibrios preferentially adhere to GM3/GM4 having monounsaturated fatty acids (2010)
  • Author(s): Matsunaga N, et al.
  • Organizer: International Carbohydrate Symposium (ICS)
  • Place of Presentation: Makuhari, Chiba, Japan
  • Year and Date: 2010-08-05
• [Presentation] 真菌グルコシルセラミド合成の編集を担う酵素EGCrP (2010)
  • Author(s): 伊東信
  • Organizer: スフィンゴリピドテラピーカンファレンス(STC)
  • Place of Presentation: 鳥取
  • Year and Date: 2010-07-16
• [Presentation] Structure and functions of sphingolipid-metabolizing enzymes (2010)
  • Author(s): Ito M
  • Organizer: Gordon Research Conference
  • Place of Presentation: Ventura, California, USA
  • Year and Date: 2010-02-10
• [Presentation] Structure and functions of sphingolipid-metabolizing machinery (2010)
  • Author(s): M.Ito
  • Organizer: Gordon Research Conference
  • Place of Presentation: Ventula, LA, USA
  • Year and Date: 2010-02-09
• [Presentation] The Crystal structure of sphingolipid ceramide N-deacylase that catalyzes the cleavage and creation of an N-acyl linkage of various glycosphingolipids (2010)
  • Author(s): & Okino N, Kakuta Y, Kato-Unoki Y, Sueyoshi N, Ito M
  • Organizer: 25th International Carbohydrate Symposium
  • Place of Presentation: Makuhari Messe International Convention Complex, Chiba, Japan
• [Presentation] Endoglycoceramidase-related protein(EGrP) is a novel glucosylceramidase that participates in glucosylceramide metabolism in pathogenic fungi (2010)
  • Author(s): Ishibashi Y, Okino N, Ito M
  • Organizer: 25th International Carbohydrate Symposium ; ICS2010
  • Place of Presentation: Makuhari Messe International Convention Complex, Chiba, Japan
• [Presentation] 真菌のGlcCer代謝を担う真菌特異的グルコシルセラミダーゼEGrP (2010)
  • Author(s): 石橋洋平、沖野望、池田和貴、田口良、伊東信
  • Organizer: 第27回内藤コンファレンス生体膜ダイナミクスと脂質生物学
  • Place of Presentation: シャトレーゼガトーキングダムサッポロ、札幌市
• [Presentation] Rhodococcus erythropolisを宿主としたエンドグリコセラミダーゼIIおよびその活性化タンパク質の発現と機能の解析 (2010)
  • Author(s): 坂口圭史、田村具博、田中勲、沖野望、伊東信
  • Organizer: 第52回日本脂質生化学会
  • Place of Presentation: 伊香保温泉、群馬県
• [Presentation] スフィンゴ脂質セラミドN-デアシラーゼ(SCDase)のX線結晶構造解析 (2009)
  • Author(s): 沖野望,角田佳充,鵜木(加藤)陽子,末吉紀行,伊東信
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド、神戸
• [Presentation] 病原性真菌より見出された新規グルコシルセラミド分解酵素 (2009)
  • Author(s): 石橋洋平,沖野望,伊東信
  • Organizer: 第82回日本生化学会大会
  • Place of Presentation: 神戸ポートアイランド、神戸
• [Presentation] ゼブラフィッシュを用いた糖脂質研究のインパクト (2009)
  • Author(s): 伊東信、座間宏太、苣田慎一、吉村征浩、松永尚之、清水耕平、沖野望、上村聡、郷信二、池田和樹、田口良、井ノ口仁一、平林義雄
  • Organizer: 第29回日本糖質学会年会
  • Place of Presentation: 高山
• [Presentation] 立体構造モデルに基づくエンドグリコセラミダーゼの特異性の解析と改変 (2009)
  • Author(s): 石橋洋平、小林宇太郎、坂口圭史、土方敦司、沖野望、伊東信
  • Organizer: 第29回日本糖質学会年会
  • Place of Presentation: 高山
• [Presentation] 接合菌Rhizopus oryzaeより発見された新規グルコシルセラミド分解酵素 (2009)
  • Author(s): 石橋洋平, 他
  • Organizer: 平成21年度日本生化学会九州支部例会
  • Place of Presentation: 福岡 九州大学大学馬出キャンパス
• [Presentation] ラビリンチュラ由来のリゾリン脂質アシル転移酵素の解析 (2009)
  • Author(s): 安部英理子, 他
  • Organizer: 平成21年度日本生化学会九州支部例会
  • Place of Presentation: 福岡 九州大学大学馬出キャンパス
• [Presentation] セラミド代謝の生物学的インパクト (2009)
  • Author(s): 伊東信
  • Organizer: 第4回スフィンゴテラピィ研究会
  • Place of Presentation: 鳥取 大山ロイヤルホテル
• [Presentation] Transglycosylation-based Fluorescent Labeling of Glycolipids by EGALC (2009)
  • Author(s): Ishibashi
  • Organizer: 50th International Conference on the Bioscience of Lipids
  • Place of Presentation: Regensburg、Germany
• [Presentation] Transglycosylation-based fluorescent labeling of glycolipids by EGALC (2007)
  • Author(s): Ishibashi Y, Nagamatsu Y, Meyer S, Okino N, Geyer G, Ito M
  • Organizer: 50th International Conference on the Bioscience of Lipids
  • Place of Presentation: Regensburg、Germany
• [Book] セタミド:基礎戸応用(セラミドの代謝)
• [Remarks]
  • URL: http://www.agr.kyushu-u.ac.jp/lab/kaishika/institute/2010insti.html
• [Patent(Industrial Property Rights)] 新規エンドグリコセラミダーゼ (1999)
  • Inventor(s): 伊東信、石橋洋平、沖野望
  • Industrial Property Rights Holder: 国立大学法人九州大学
  • Acquisition Date: 1999-08-14
• [Patent(Industrial Property Rights)] 新規タンパク質 (1999)
  • Inventor(s): 大森.、伊東信
  • Industrial Property Rights Holder: 三菱化学株式会社
  • Acquisition Date: 1999-10-09