Elucidation of biological function of glycan by integrated omics analysis based on glycoproteome.

• Project/Area Number: 15H01841
• Research Category: Grant-in-Aid for Scientific Research (A)
• Allocation Type: Single-year Grants
• Section: 一般
• Research Field: Chemical biology
• Research Institution: National Institute of Advanced Industrial Science and Technology
• Principal Investigator: Narimatsu Hisashi 国立研究開発法人産業技術総合研究所, 生命工学領域, 招聘研究員 (40129581)
• Co-Investigator(Kenkyū-buntansha): 梶 裕之 国立研究開発法人産業技術総合研究所, .生命工学領域, 研究チーム長 (80214302) ; 栂谷内 晶 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (60392635) ; 佐藤 隆 国立研究開発法人産業技術総合研究所, 生命工学領域, 主任研究員 (90371046) ; 安形 清彦 国立研究開発法人産業技術総合研究所, 生命工学領域, 招聘研究員 (00611138)
• Project Period (FY): 2015-04-01 – 2018-03-31
• Project Status: Completed (Fiscal Year 2017)
• Budget Amount: ¥40,300,000 (Direct Cost: ¥31,000,000、Indirect Cost: ¥9,300,000); Fiscal Year 2017: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2016: ¥11,700,000 (Direct Cost: ¥9,000,000、Indirect Cost: ¥2,700,000); Fiscal Year 2015: ¥16,900,000 (Direct Cost: ¥13,000,000、Indirect Cost: ¥3,900,000)
• Keywords: 糖鎖 / 糖転移酵素 / 糖タンパク質 / 糖ペプチド / 糖鎖機能 / グライコプロテオミクス / グライコミクス / 糖鎖改変 / グライコーム解析 / プロテオーム / 疾患モデル / トランスクリプトーム / グライコプロテオーム解析 / トランスクリプトーム解析

Outline of Final Research Achievements

One of the methods to elucidate the biological function of glycan is to modify the expression of glycosyltransferases responsible for the biosynthesis of specific glycans and to analyze the phenotypes caused thereby. Moreover, to elucidate the relationship between glycan and biological function, glycosylated sites, glycan variation (glycome), and its proportions for each glycoprotein must be analyzed, if the determined glycan-carriers are of interest. Therefore, we developed a novel glycoproteomics technology to reveal the site-specific glycome in several glycan-modified cells/mice. As a result, we identified many carrier glycoproteins of the glycosyltransferase-specific glycans, including Lewis x, LacdiNAc, pLN and chondroitin sulfate structures, and tried to elucidate the molecules related to the phenotype and its molecular mechanism. Our novel glycoproteomics method provided important insights into the biological function of glycans.

Research Products

• [Journal Article] Incomplete clearance of apoptotic cells by core 1-derived O-glycan-deficient resident peritoneal macrophages. (2018)
  • Author(s): Wakui H, Fuseya S, Suzuki R, Shimbo M, Okada R, Hamada M, Kuno A, Hagiwara K, Sato T, Narimatsu H, Kudo T, Takahashi S.
  • Journal Title: Biochem Biophys Res Commun Volume: 122 Issue: 2 Pages: 2017-2023
  • DOI: 10.1016/j.bbrc.2017.12.066
  • Peer Reviewed
• [Journal Article] Identification of Poly-N-Acetyllactosamine-Carrying Glycoproteins from HL-60 Human Promyelocytic Leukemia Cells Using a Site-Specific Glycome Analysis Method, Glyco-RIDGE. (2018)
  • Author(s): Togayachi A, Tomioka A, Fujita M, Sukegawa M, Noro E, Takakura D, Miyazaki M, Shikanai T, Narimatsu H, Kaji H.
  • Journal Title: J Am Soc Mass Spectrom. Volume: - Issue: 6 Pages: 1-15
  • DOI: 10.1007/s13361-018-1938-6
  • Peer Reviewed / Open Access
• [Journal Article] Postnatal lethality and chondrodysplasia in mice lacking both chondroitin sulfate N-acetylgalactosaminyltransferase-1 and -2. (2017)
  • Author(s): Shimbo M, Suzuki R, Fuseya S, Sato T, Kiyohara K, Hagiwara K, Okada R, Wakui H, Tsunakawa Y, Watanabe H, Kimata K, Narimatsu H, Kudo T, Takahashi S.
  • Journal Title: PLoS One. Volume: 12 Issue: 12 Pages: e0190333-e0190333
  • DOI: 10.1371/journal.pone.0190333
  • NAID: 120007134461
  • Peer Reviewed / Open Access
• [Journal Article] Contribution of Lewis X Carbohydrate Structure to Neuropathogenic Murine Coronaviral Spread (2016)
  • Author(s): Kakizaki M, Togayachi A, Narimatsu H, Watanabe R.
  • Journal Title: Japanese Journal of Infectious Diseases Volume: 69 Issue: 5 Pages: 405-413
  • DOI: 10.7883/yoken.JJID.2015.499
  • NAID: 130005416321
  • ISSN: 1344-6304, 1884-2836
  • Peer Reviewed
• [Journal Article] Formation of fibroblastic reticular network in the brain after infection with neurovirulent murine coronavirus (2016)
  • Author(s): R. Watanabe, M. Kakizaki, Y. Ikehara, A. Togayachi
  • Journal Title: Neuropathology Volume: 36（6） Issue: 6 Pages: 513-526
  • DOI: 10.1111/neup.12302
  • Peer Reviewed
• [Journal Article] Large-scale mutational analysis in the EXT1 and EXT2 genes for Japanese patients with multiple osteochondromas. (2016)
  • Author(s): Ishimaru D, Gotoh M, Takayama S, Kosaki R, Matsumoto Y, Narimatsu H, Sato T, Kimata K, Akiyama H, Shimizu K, Matsumoto K.
  • Journal Title: BMC Genet. Volume: 17 Issue: 1 Pages: 52-52
  • DOI: 10.1186/s12863-016-0359-4
  • Peer Reviewed / Open Access
• [Journal Article] Comparison of analytical methods for profiling N- and O-linked glycans from cultured cell lines : HUPO Human Disease Glycomics/Proteome Initiative multi-institutional study. (2015)
  • Author(s): Ito H, Kaji H, Togayachi A, Narimatsu H.et al.
  • Journal Title: Glycoconj J. Volume: なし
  • Peer Reviewed / Int'l Joint Research
• [Journal Article] Large-Scale Identification of N-Glycan Glycoproteins Carrying Lewis x and Site-Specific N-Glycan Alterations in Fut9 Knockout Mice. (2015)
  • Author(s): Noro E, Togayachi A, Sato T, Tomioka A, Fujita M, Sukegawa M, Suzuki N, Kaji H, Narimatsu H.
  • Journal Title: J Proteome Res. Volume: 14(9) Issue: 9 Pages: 3823-3834
  • DOI: 10.1021/acs.jproteome.5b00178
  • Peer Reviewed
• [Journal Article] Large-scale identification of secretome glycoproteins recognized by Wisteria floribunda agglutinin: A glycoproteomic approach to biomarker discovery. (2015)
  • Author(s): Sugahara D, Tomioka A, Sato T, Narimatsu H, Kaji H.
  • Journal Title: Proteomics. Volume: 15(17) Issue: 17 Pages: 2921-33
  • DOI: 10.1002/pmic.201400443
  • Peer Reviewed
• [Journal Article] Influenza A Virus-Induced Expression of a GalNAc Transferase, GALNT3, via MicroRNAs Is Required for Enhanced Viral Replication. (2015)
  • Author(s): Nakamura S, Horie M, Daidoji T, Honda T, Yasugi M, Kuno A, Komori T, Okuzaki D, Narimatsu H, Nakaya T, Tomonaga K.
  • Journal Title: J Virol. Volume: 90 Issue: 4 Pages: 1788-801
  • DOI: 10.1128/jvi.02246-15
  • Peer Reviewed / Open Access
• [Presentation] Fut9ノックアウトマウスを用いた脳におけるルイスｘキャリアタンパク質の網羅的な解析（Large-scale identification of glycoprotein carrying Lewis x in the brain using Fut9 knockout mice） (2017)
  • Author(s): 野呂絵里花、栂谷内晶、富岡あづさ、助川昌子、藤田弥佳、鈴木奈美、梶裕之、成松久
  • Organizer: 日本糖質学会
• [Presentation] Deletion of B3glct disrupts craniofacial, skeletal, and cardiac development in mice (2017)
  • Author(s): Haltiwanger R, Rubel D, Grady R, Uddin S, Komatsu D, Bamforth S, Schneider J, Sato T, Narimatsu H, Holdener B.
  • Organizer: 24th INTERNATIONAL SYMPOSIUM ON GLYCOCONJUGATES（Jeju, Korea）
  • Int'l Joint Research
• [Presentation] マウス卵管におけるフコシル化糖鎖sialyl Lewis Xの生合成経路の解析 (2017)
  • Author(s): 佐藤裕之、松浦寛明、小林純子、栂谷内晶、成松久、Kay-Hooi Khoo、川島博人
  • Organizer: 第61回日本薬学会関東支部大会
• [Presentation] Developement of site-specific glycome analysis method of complex glycoproteome (2017)
  • Author(s): Azusa Tomioka, Chiaki Okatani, Erika Noro, Toshihide Shikanai, Hisashi Narimatsu. Hiroyuki Kaji
  • Organizer: HUPO World Congress （Dublin, Ireland）
  • Int'l Joint Research
• [Presentation] Identification of glycoproteins with novel LacdiNAc structures synthesized by B3GALNT2. (2017)
  • Author(s): Takahiro Nakane, Kiyohiko Angata, Takashi Sato, Hiroyuki Kaji, and Hisashi Narimatsu
  • Organizer: The 9th ACGG conference（Hong Kong, China)
  • Int'l Joint Research
• [Presentation] β3GalNAcT2糖転移酵素はtype I LacdiNAc構造をN型糖鎖とO型糖鎖上に形成する (2017)
  • Author(s): 安形清彦、中根隆浩、古川早苗、佐藤隆、澤木弘道、曽我部万紀、久保田智己、梶裕之、成松久
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] β3GalNT2糖転移酵素によって合成されるtype I LacdiNAc構造キャリアタンパク質の同定 (2017)
  • Author(s): 中根隆浩、安形清彦、佐藤隆、梶裕之、成松久
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] 腎糸球体足細胞におけるムチン型O結合型糖鎖の機能解明 (2017)
  • Author(s): 布施谷清香, 岡田理沙, 鈴木陸, 新保未来, 涌井宏優, 中村勇輝, 坪内鴻奈, 臼井俊明, 森戸直記, 萩原梢, 佐藤隆, 成松久, 高橋智, 工藤崇
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] マクロファージでのTim4タンパク質の安定発現にムチン型糖鎖は必須である (2017)
  • Author(s): 涌井宏優, 布施谷清香, 岡田理沙, 鈴木陸, 新保未来, 濱田理人, 萩原梢, 佐藤隆, 成松久, 高橋智, 工藤崇
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] 全身性誘導コンディショナルノックアウトマウスを用いたムチン型糖鎖新規機能 (2017)
  • Author(s): 鈴木陸, 中村勇輝, 涌井宏優, 岡田理沙, 布施谷清香, 坪内鴻奈, 萩原梢, 佐藤隆, 成松久, 高橋智, 工藤崇
  • Organizer: 2017年度生命科学系学会合同年次大会
• [Presentation] 複雑性の極み： 糖鎖修飾状態の解析への挑戦 (2017)
  • Author(s): 梶裕之、富岡あづさ、野呂絵里花、岡谷千晶、藤田弥佳、助川昌子、成松久
  • Organizer: 2017年度生命科学系学会合同年次大会
  • Invited
• [Presentation] B3GALNT2糖転移酵素によって合成される新規LacdiNAc構造キャリアタンパク質の同定 (2016)
  • Author(s): 中根 隆浩, 安形 清彦, 佐藤 隆, 梶 裕之, 成松 久
  • Organizer: 第３９回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-12-01
• [Presentation] 糖タンパク質大規模解析とデータベース構築： 糖鎖付加位置マッピングから位置ごとのグライコーム解析へ (2016)
  • Author(s): 梶 裕之, 富岡あづさ, 野呂絵里花, 藤田弥佳, 助川昌子, 岡谷千晶, 鹿内俊秀, 成松久
  • Organizer: 第３９回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-30
• [Presentation] Both Chondroitin Sulfate N-Acetylgalactosaminyltransferase-1 and -2 are Required for Endochondral Ossification (2016)
  • Author(s): Miki Shimbo, Risa Okada, Hiromasa Wakui, Riku Suzuki, Sayaka Fuseya, Kozue Hagiwara, Takashi Sato, Hisashi Narimatsu, Satoru Takahashi, and Takashi Kudo.
  • Organizer: 第３９回日本分子生物学会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-28
• [Presentation] 腎糸球体足細胞におけるムチン型糖鎖の役割 (2016)
  • Author(s): 布施谷清香, 岡田理沙, 鈴木陸, 新保未来, 涌井宏優, 臼井俊明, 森戸直記, 萩原梢, 佐藤隆, 成松久, 高橋智, 工藤崇
  • Organizer: 第３９回日本分子生物学会年会
  • Place of Presentation: パシフィコ横浜（神奈川県横浜市）
  • Year and Date: 2016-11-28
• [Presentation] Successful translational research based on combination of multiple glyco- proteomics technologies. (2016)
  • Author(s): 成松久
  • Organizer: Swedish MS/proteomic meeting
  • Place of Presentation: University of Gothenburg(Gothenburg,Sweden)
  • Year and Date: 2016-11-20
  • Int'l Joint Research / Invited
• [Presentation] 最近のグライコプロテオミクス： 糖鎖付加部位ごとの糖鎖不均一性の大規模解析法の開発 (2016)
  • Author(s): 梶 裕之
  • Organizer: 第14回日本糖鎖科学コンソーシアム(JCGG)シンポジウム
  • Place of Presentation: 御茶ノ水ソラシティカンファレンスセンター（東京都千代田区）
  • Year and Date: 2016-11-01
• [Presentation] 佐藤隆, 梶裕之, 千葉靖典, 成松久 (2016)
  • Author(s): Characterization of Wisteria Floribunda lectin and Its Application for Glycoproteomics Study
  • Organizer: 8th Asian Community of Glycoscience and Glycotechnology
  • Place of Presentation: Jiangnan University (Wuxi, China)
  • Year and Date: 2016-10-14
  • Int'l Joint Research / Invited
• [Presentation] Analyses of glycogene expression by RNA sequencing to determine the different glycan synthesis. (2016)
  • Author(s): 安形清彦, 辻川紫華子, 佐藤隆, 栂谷内晶, 成松久
  • Organizer: 8th Asian Community of Glycoscience and Glycotechnology
  • Place of Presentation: Jiangnan University (Wuxi, China)
  • Year and Date: 2016-10-14
  • Int'l Joint Research / Invited
• [Presentation] 糖タンパク質の糖鎖不均一性の大規模解析 (2016)
  • Author(s): 梶 裕之, 富岡あづさ, 野呂絵里花, 藤田弥佳, 助川昌子, 岡谷千晶, 鹿内俊秀, 成松久
  • Organizer: 第89回日本生化学会大会
  • Place of Presentation: 仙台国際センター／東北大学川内北キャンパス（宮城県仙台市）
  • Year and Date: 2016-09-27
• [Presentation] Accurate mass- and glycan composition-based assignment of glycosylation site-specific glycomes of complex glycoprotein mixture (2016)
  • Author(s): 梶 裕之, 富岡あづさ, 野呂絵里花, 藤田弥佳, 助川昌子, 岡谷千晶, 鹿内俊秀, 成松久,
  • Organizer: Human Proteome Organization World Congress
  • Place of Presentation: Taipei International Convention Center（Taipei,Taiwan)
  • Year and Date: 2016-09-20
  • Int'l Joint Research / Invited
• [Presentation] Use of whole transcriptome analysis to determine glycogene expression in hepatic cells (2016)
  • Author(s): 安形清彦, 澤木弘道, 辻川紫華子, 雄長誠, 栂谷内晶, 成松久
  • Organizer: 6th Charles Waren Workshop 2016
  • Place of Presentation: 北海道大学（北海道札幌市）
  • Year and Date: 2016-08-24
  • Int'l Joint Research
• [Presentation] Large-scale identification of glycoproteins carrying Lewis x and site-specific glycan alterations in Fut9 knockout mice. (2016)
  • Author(s): Erika Noro, Hiroyuki Kaji, Hisashi Narimatsu.
  • Organizer: KHUPO 16th Annual International Proteomics Conference
  • Place of Presentation: Daejeon Convention Center（Daejeon, Korea）
  • Year and Date: 2016-03-28
  • Int'l Joint Research / Invited
• [Presentation] Roles of Chondroitin Sulfate N-acetylgalactosaminyltransferase-1 and -2 in Endochondral Ossification. (2015)
  • Author(s): Miki Shimbo, Takashi Sato, Kozue Hagiwara, Risa Okada, Hiromasa Wakui, Hisashi Narimatsu, Satoru Takahashi, and Takashi Kudo.
  • Organizer: BMB2015
  • Place of Presentation: 神戸ポートアイランド（兵庫県神戸市）
  • Year and Date: 2015-12-01
• [Presentation] Novel LacdiNAc structures on N- and O-glycans synthesized in vivo by 3GalNAcT2. (2015)
  • Author(s): Kiyohiko Angata, Takahiro Nakane, Sanae Furukawa, Takashi Sato, Hiromichi Sawaki, Maki Sogabe, Tomomi Kubota, Hiroyuki Kaji, and Hisashi Narimatsu.
  • Organizer: The 7th ACGG conference
  • Place of Presentation: ホテル大観荘（宮城県松島市）
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] Identification of glycoproteins with novel LacdiNAc structures synthesized by B3GALNT2. (2015)
  • Author(s): Takahiro Nakane, Kiyohiko Angata, Takashi Sato, Hiroyuki Kaji, and Hisashi Narimatsu
  • Organizer: The 7th ACGG conference
  • Place of Presentation: ホテル大観荘（宮城県松島市）
  • Year and Date: 2015-11-13
  • Int'l Joint Research
• [Presentation] α1,3-フコース転移酵素9ノックアウトマウスを用いた新規グライコプロテオミクス技術による糖鎖付加部位特異的グライコームの網羅的な解析 (2015)
  • Author(s): 野呂絵里花、栂谷内晶、佐藤隆、富岡あづさ、藤田弥佳、助川昌子、鈴木奈美、梶裕之、成松久
  • Organizer: 第34回日本糖質学会
  • Place of Presentation: 東京大学安田講堂（東京都文京区）
  • Year and Date: 2015-07-31
• [Presentation] α1,3-フコース転移酵素9ノックアウトマウスを用いた糖鎖付加部位特異的グライコームの網羅的な解析 (2015)
  • Author(s): 野呂絵里花、栂谷内晶、佐藤 隆、富岡 あづさ、藤田弥佳、助川昌子、鈴木奈美、梶裕之、成松久
  • Organizer: JHUPO第13回大会
  • Place of Presentation: くまもと森都心プラザ（熊本県熊本市）
  • Year and Date: 2015-07-23
• [Book] A Practical Guide to Using Glycomics Databases：GlycoGene Database (GGDB) on the Semantic Web. (2016)
  • Author(s): 成松久, 鈴木芳典, 木下フローラ聖子, 鹿内俊秀, 藤田典昭, 佐藤隆, 栂谷内晶, 横尾岳彦, 安形清彦, 久保田智巳, 野呂絵里花
  • Total Pages: 13
  • Publisher: Springer