リーダー 水野　真盛 メンバー 大隅　賢二 黒河内　政樹 後藤　浩太朗 筒井　正斗 八須　和子 弘瀬　友理子 （五十音順） 学生 学部4年生　3名（東海大学２名、北里大学１名）

　糖鎖は核酸やタンパク質と同様に、天然に広く存在する主要な生体物質であります。糖鎖は主にタンパク質や脂質と結合した「糖タンパク質」や「糖脂質」のような「複合糖質」として生体内に存在しています。糖鎖の機能の一つとして認識素子（＝目印）としての働きが知られています。例えば、赤血球上の糖鎖構造の違いによって血液型が決定されます（血液型の目印）。また、細胞上の特定の糖鎖を目指してウイルスや毒素は細胞に接近し感染します（感染の目印）。最近では病気になると体内の糖鎖構造が変化することが知られており、この変化した糖鎖を調べる事で病気の診断を行う研究もされています（病気の目印）。これら以外にも、我々の体を病原菌やウイルスから守ってくれる「抗体」や赤血球の生産に欠かせないエリスロポエチンは糖タンパク質であり、これらから糖鎖を除去してしまうと正しく機能しなくなってしまうことが知られています。このように糖鎖は我々の生命現象に非常に深く係わっている重要な物質であります。
　当研究室では糖・複合糖質・糖タンパク質合成に向けた、有機合成・酵素合成の要素技術の開発に関する研究、及び糖・糖タンパク質糖鎖の質量分析による構造解析に関する研究を行っています。

【合成研究】
　糖と糖を結合させた糖鎖、または糖をタンパク質や脂質に結合させた複合糖質の合成、糖を原料とした有用物質の合成、及びこれら糖誘導体の迅速・簡便な新規合成手法の開発を手がけています。これまでに、化学－酵素法による糖ペプチドの合成、糖と糖をつなぐグリコシル化反応の開発、糖とアミノ酸との結合構築法の開発、糖ペプチドの機能解明、化学－酵素法を用いた複雑な構造を有する天然型糖鎖の合成などを行ってきました。最近は、糖を足場としたタンパク質の位置選択的な修飾化法の開発も行っています。またこの技術を活用して、抗体分子の特定部位に特定数の薬物を搭載した抗体薬物複合体（Antibody Drug Conjugate：ADC）の合成に関する研究も行っています。

【構造解析研究】
　糖鎖は分岐構造を有するため、遺伝子やタンパク質と比べると複雑な構造をしています。現在糖鎖の構造解析は、使用するサンプル量の少なさや測定の簡便さなどから質量分析装置による解析が主流となっています。当研究室では診断の際のバイオマーカーとなる糖鎖構造を明らかにすることを目的に、質量分析法による糖タンパク質の分析技術開発を行っています。

現在取り組んでいる研究テーマは以下のとおりです。
メインテーマ
「糖質科学の研究進展に寄与する新規技術の開発、及び探索研究」

サブテーマ
(1) 薬物の搭載部位及び搭載数の制御が可能な抗体薬物複合体（ADC）の合成法の開発
(2) 質量分析法を用いた糖鎖構造の新規解析法の開発
(3) グライコプロテオミクス解析手法の確立を目指した技術開発
(4) 生化学的・医化学的知見に有用な糖質誘導体の合成と機能解明
(5) 糖タンパク質、及び糖ペプチドの効率的合成法の開発

２０２０年

The GlyCosmos Portal: a unified and comprehensive web resource for the glycosciences. Issaku Yamada, Masaaki Shiota, Daisuke Shinmachi, Tamiko Ono, Shinichiro Tsuchiya, Masae Hosoda, Akihiro Fujita, Nobuyuki P. Aoki, Yu Watanabe, Noriaki Fujita, Kiyohiko Angata, Hiroyuki Kaji, Hisashi Narimatsu, Shujiro Okuda & Kiyoko F. Aoki-Kinoshita Nature Methods 2020, 17, 649–650 https://www.nature.com/articles/s41592-020-0879-8

Preparation of O-Glycopeptides from commercial bovine whey proteins using offline liquid chromatography–Mass spectrometry Masaki Kurogochi, Akio Matsuda, MamoruMizuno Carbohydr. Res., 2020, 491, 107981 https://doi.org/10.1016/j.carres.2020.107981

Site-specific protein PEGylation catalyzed by endo-β-N-acetylglucosaminidase Kohtaro Goto, Masaki Kurogochi, Shou Takashima, Masako Mori, Akio Matsuda, and Mamoru Mizuno* Tetrahedron Lett., 2020, 61, 151475 https://doi.org/10.1016/j.tetlet.2019.151475

Crystal structures of fukutin-related protein (FKRP), a ribitol-phosphate transferase related to muscular dystrophy Naoyuki Kuwabara, Rieko Imae, Hiroshi Manya, Tomohiro Tanaka, Mamoru Mizuno, Hiroki Tsumoto, Motoi Kanagawa, Kazuhiro Kobayashi, Tatsushi Toda, Toshiya Senda, Tamao Endo, and Ryuichi Kato Nat. Commun., 2020, 11, Article number 303 https://doi.org/10.1038/s41467-019-14220-z

２０１９年

Efficient Synthesis of N- and O-linked Glycopeptides using Acid-Labile Boc Groups for the Protection of Carbohydrate Moieties Tomohiro Tanaka, Mika Shiraishi, Akio Matsuda and Mamoru Mizuno* Tetrahedron Lett., 2019, 60, 151106

２０１８年

Glycan Remodeling of Glycoproteins Using ENGases Masaki Kurogochi Trends in Glycoscience and Glycotechnology, 2018, 30, E209-E219

GlycanFormatConverter: a conversion tool for　translating the complexities of glycans Shinichiro Tsuchiya, Issaku Yamada and Kiyoko F. Aoki-Kinoshita Bioinformatics, 2018, 1–7

Meeting Report of the International Life Science Integration Workshop 2018 Issaku Yamada、他33名 Glycobiology , 2018, 28, 552 – 555

Synthesis of 1,5-Anhydro-D-fructose derivatives and evaluation of their inflammasomeinhibitors Kohtaro Goto, Hiroko Ideo, Akiko Tsuchida, Yuriko Hirose, Ikuro Maruyama, Satoshi Noma, Takashi Shirai, Junko Amano, Mamoru Mizuno, Akio Matsuda Bioorg. Med. Chem. 2018, 26, 3763-3772

Cell endogenous activities of fukutin and FKRP coexist with TMEM5 Ryuta Nishihara, Kazuhiro Kobayashi, Reiko Imae, Hiroki Tsumoto, Hiroshi Manya, Mamoru Mizuno, Motoi Kanagawa, Tamao Endo, and Tatsushi Toda Biochemical and Biophysical Research Communications 2018, 497, 1025-1030.

CDP-glycerol inhibits the synthesis of the functional O-mannosyl glycan of α-dystroglycan Rieko Imae, Hiroshi Manya, Hiroki Tsumoto, Kenji Osumi, Tomohiro Tanaka, Mamoru Mizuno, Motoi Kanagawa, Kazuhiro Kobayashi, Tatsushi Toda, Tamao Endo J. Boil. Chem., 2018, 293, 12186-12198

Binding Assessment of Monosaccharide-Boronic Acid Complexes via Tandem Mass Spectrometry Masaki Kurogoch, Mamoru Mizuno and Akio Matsuda ChemistrySelect, 2018, 3, 8193-8198

２０１７年

3D structural analysis of Protein O-Mannosyl Kinase POMK, a causative geneproduct of dystroglycanopathy Masamichi Nagae, Sushil K. Mishra, Makiko Neyazaki, Rika Oib, Akemi Ikeda, Naohiro Matsugaki, Satoko Akashi, Hiroshi Manya, Mamoru Mizuno, Hirokazu Yagi, Koichi Kato, Toshiya Senda, Tamao Endo, Terukazu Nogi and Yoshiki Yamaguchi GENES CELLS 2017, 22, 348-359

Isolation of a methylated mannose-binding protein　from terrestrial worm Enchytraeus japonensis Shigeru Ogawa, Mamoru Mizuno, Mayuko Suzuki, Kohtaro Goto, Yuriko Hirose, Akio Matsuda, Takuya Saito, Suguru Oguri and Kiyoshi Furukawa Glycoconjugate J., 2017, 34(5), 591-601

フルオラス合成法の糖質合成への展開 水野真盛 有機合成化学協会誌（2017） vol.75 No.6, p622

２０１６年

Identification of a Post-translational Modification with Ribitol-Phosphate and Its Defect in Muscular Dystrophy Motoi Kanagawa, Kazuhiro Kobayashi, Atsushi Kuga, Michiko Tajiri, Hiroshi Manya, Yoshiki Yamaguchi, Keiko Akasaka-Manya, Jun-ichi Furukawa, Mamoru Mizuno, Hiroko Kawakami, Yasuro Shinohara, Yoshinao Wada, Tamao Endo, Tatsushi Toda Cell Reports 2016, 14, 2209-2223

Carbohydrate-binding domain of the POMGnT1 stem region modulates O-mannosylation sites of α-dystroglycan Naoyuki Kuwabara, Hiroshi Manya, Takeyuki Yamada, Hiroaki Tateno, Motoi Kanagawa, Kazuhiro Kobayashi, Keiko Akasaka-Manya, Yuriko Hirose, Mamoru Mizuno, Mitsunori Ikeguchi, Tatsushi Toda, Jun Hirabayashi, Toshiya Senda, Tamao Endo, Ryuichi Kato Proc. Natl. Acad. Sci. (PNAS) 2016, 113, 9280-9285

The Muscular Dystrophy Gene TMEM5 Encodes a Ribitol β1-4 Xylosyltransferase Required for the Functional Glycosylation of Dystroglycan Hiroshi Manya, Yoshiki Yamaguchi, Motoi Kanagawa, Kazuhiro Kobayashi, Michiko Tajiri, Keiko Akasaka-Manya, Hiroko Kawakami, Mamoru Mizuno, Yoshinao Wada, Tatsushi Toda, and Tamao Endo J. Biol. Chem., 2016, 291, 24618-24627

２０１５年

Glycoengineered Monoclonal Antibodies with Homogeneous Glycan (M3, G0, G2, and A2) using a chemoenzymatic approach have different affinities for FcγRIIIa and variable antibody-dependent cellular cytotoxicity activity

Masaki Kurogochi, Masako Mori, Kenji Osumi, Mami Tojino, Shu-ichi Sugawara, ShouTakashima, Yuriko Hirose, Wataru Tsukimura, Mamoru Mizuno, Junko Amano, Akio Matsuda, Masahiro Tomita, Atsushi Takayanag5, Shin-Ichiro Shoda, Takashi Shirai PLOS ONE, DOI:10.1371/journal.pone.0132848

Kazuo Fukuda, Mami Tojino, Kohtaro Goto, Hirofumi Dohi, Yoshihiro Nishida, Mamoru Mizuno "A recyclable heavy fluorous tag carrying an allyl alcohol pendant group: design and evaluation toward applications in synthetic carbohydrate chemistry" Carbohydr. Res., 2015, 407, 122-130

２０１４年

Mamoru Mizuno "Carbohydrate-Lectin Interaction Assay by Fluorescence Correlation Spectroscopy Using Fluorescence-Labeled Glycosylasparagines" Lectins, Methods Mol Biol. 2014, 1200, 215-221. doi: 10.1007/978-1-4939-1292-6_19.

Kazuo Fukuda, Mami Tojino, Kohtaro Goto, Hirofumi Dohi, Yoshihiro Nishida, Mamoru Mizuno "Preparation of acid-resistant heavy fluorous tags for recycling in synthetic systems" J. Fluorine Chem. 2014, 166, 52-59 doi:10.1016/j.jfluchem.2014.07.014

Katalin Uray, Mamoru Mizuno, Toshiyuki Inazu, Kohtaro Goto, Zsuzsa Majer, Ferenc Hudecz "The effect of glycosylation on the antibody recognition of a MUC2 mucin epitope" Biopolymers (Peptide Science), 2014, 102, 390-395 doi:10.1002/bip.22526

T. Katayama, I. Yamada, T. Takagi, et al. "BioHackathon series in 2011 and 2012: penetration of ontology and linked data in life science domains." J. Biomed. Semantics. 2014, 5. doi:10.1186/2041-1480-5-5

K. Takana, I. Yamada, H. Narimatsu, et al., "WURCS: the Web3 unique representation of carbohydrate structures" J. Chem. Inf. Model. 2014, 54, 1558-66. doi: 10.1021/ci400571e

R. Ranzinger, I. Yamada, H. Narimatsu, et al., "GlycoRDF: an ontology to standardize glycomics data in RDF" Bioinformatics. 2014 Nov 11. pii: btu732. doi: 10.1093/bioinformatics/btu732

２０１３年

Mami Tojino, Yuriko Hirose and Mamoru Mizuno "Convenient Synthesis of Glycosyl Bromide from 1-O-acetyl Sugars by Photo-irradiative Phase-Vanishing Reaction of Molecular Bromine" Tetrahedron Lett. 2013, 54, 7124-7126 doi:10.1016/j.tetlet.2013.10.088

Goto, Kohtaro; and Mizuno, Mamoru "Application of Fluorous Chemistry for Oligosaccharide Synthesis" Trends in Glycoscience and Glycotechnology, 2013, 25, 203-213. doi:10.4052/tigg.25.203

Kiyoko F Aoki-Kinoshita, Issaku Yamada, Hisashi Narimatsu, et al., "Introducing glycomics data into the Semantic Web" Journal of Biomedical Semantics 2013, 4(39). doi:10.1186/2041-1480-4-39

Kiyoko F Aoki-Kinoshita, Issaku Yamada, Hisashi Narimatsu, et al., "The Third ACGG-DB Meeting Report: Towards an international collaborative infrastructure for glycobioinformatics" Glycobiology 2013, 23, 144-146 doi: 10.1093/glycob/cws167

Kiyoko F Aoki-Kinoshita, Issaku Yamada, Hisashi Narimatsu, et al., "The Fifth ACGG-DB Meeting Report: Towards an International Glycan Structure Repository" Glycobiology 2013, 23, 1422-1424

２０１２年

Mami Tojino; Masako Mori; Maria Carmelita Z Kasuya; Kenichi Hatanaka; Atsushi Kawaguchi; Kyosuke Nagata; Mamoru Mizuno "Immobilization of Fluorous Oligosaccharide Recognized by Influenza Virus on Polytetrafluoroethylene Filter" Bioorg. Med. Chem. Lett., 2012, in press

Ganesh P. Subedi, Tadashi Satoh, Shinya Hanashima, Akemi Ikeda, Hiroshi Nakada, Reiko Sato, Mamoru Mizuno, Noriyuki Yuasa, Yoko Fujita-Yamaguchi and Yoshiki Yamaguchi "Overproduction of anti-Tn antibody MLS128 single-chain Fv fragment in Escherichia coli cytoplasm using a novel pCold-PDI vector" Protein Expr Purif., 2012, 82, 197-204.

Masato Noguchi, Miwa Nakamura, Ayaka Ohno, Tomonari Tanaka, Atsushi Kobayashi, Masaki Ishihara, Masaya Fujita, Akiko Tsuchida, Mamoru Mizuno and Shin-ichiro Shoda "A dimethoxytriazine type glycosyl donor enables a facile chemo-enzymatic route toward α-linked N-acetylglucosaminyl-galactose disaccharide unit from gastric mucin" Chem. Commun., 2012, 48, 5560-5562 doi: 10.1039/c2cc30946g

Yinan Wang, Tomoaki Kumazawa, Keisuke Shiba, Kenji Osumi, Mamoru Mizuno, Toshinori Sato "Glycosylation of Nα-lauryl-O-(β-D-xylopyranosyl)-L-serinamide as a Saccharide Primer in Cells" Carbohydrate. Res., 2012, 361, 33-40.

Ayano Matsumoto-Takasaki, Shinya Hanashima, Ami Aoki, Noriyuki Yuasa, Haruhiko Ogawa, Reiko Sato, Hiroko Kawakami, Mamoru Mizuno, Hiroshi Nakada, Yoshiki Yamaguchi and Yoko Fujita-Yamaguchi, "Surface plasmon resonance and NMR analyses of anti Tn-antigen MLS128 monoclonal antibody binding to two or three consecutive Tn-antigen clusters" J. Biochem. 2012, 151, 73-282 doi:10.1093/jb/mvr138

２０１１年

M. Fujita, A. Tsuchida, A. Hirata, N. Kobayashi, K. Goto, K. Osumi, Y. Hirose, J. Nakayama, T. Yamanoi, H. Ashida, and M. Mizuno "Glycoside hydrolase family 89 α-N-acetylglucosaminidase from clostridium perfringens specifically acts on GlcNAc 1,4Galβ1R at the non-reducing terminus of O-glycans in gastric mucin" J. Biol. Chem., 2011, 286, 6479-6489.

Masako Mori, Maria Carmelita Zulueta Kasuya, Mamoru Mizuno and Kenichi Hatanaka "Thiolactosides: Scaffolds for the synthesis of glycolipids in animal cells" Int. J. Carbohydr. Chem., 2011, doi:10.1155/2011/267208

Kohtaro Goto, Nueramina Nuermaimaiti, and Mamoru Mizuno* "Effective Application of Heavy Fluorous Thioglycoside for Oligosaccharide Synthesis" Chem. Lett., 2011, 40, 756-757

Keiko Akasaka-Manya, Hiroshi Manya, Mamoru Mizuno, Toshiyuki Inazu, Tamao Endo "Effects of length and amino acid sequence of O-mannosyl peptides on substrate specificity of protein O-linked mannose 1,2-N-acetylglucosaminyltransferase 1 (POMGnT1)" Biochem. Biophys. Res. Commun., 2011, 410, 632-636

２０１０年

Maria Carmelita Z. Kasuya, Mami Tojino, Mamoru Mizuno, Kenichi Hatanaka "Fluorous Tag Method for the Simultaneous Synthesis of Different Kinds of Glycolipids" Journal of Fluorine Chemistry, 131(5), 655-659, 2010/doi:10.1016/j.jfluchem.2010.02.001

Junko Amano, Takashi Nishikaze, Fumio Tougasaki, Hiroshi Jinmei, Ichiro Sugimoto,Shu-ichi Sugawara, Masaya Fujita, Kenji Osumi, Mamoru Mizuno "Derivatization with 1-Pyrenyldiazomethane Enhances Ionization of Glycopeptides but Not Peptides in Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry" Analytical Chemistry, 82(20), 8738-8743, 2010/doi:10.1021/ac101555a

Kohtaro Goto, Mamoru Mizuno "Practical Heavy Fluorous Tag for Carbohydrate Synthesis with Minimal Chromatographic Purification" Tetrahedron Letters, 51(50), 6539-6541, 2010/doi:10.1016/j.tetlet.2010.10.016

後藤浩太朗、川上宏子、水野真盛 "フルオラスケミストリーを基盤とした効率的糖鎖合成法の開発" 化学工業, 61, 186-190, 2010

Keiko Sakai, Noriyuki Yuasa, Kazuhiro Tsukamoto, Ayano Takasaki-Matsumoto, Yukiko Yajima, Reiko Sato, Hiroko Kawakami, Mamoru Mizuno, Atsushi Takayanagi, Nobuyoshi Shimizu, Munehiro Nakata and Yoko Fujita-Yamaguchi "Isolation and characterization of antibodies against three consecutive Tn-antigen clusters from a phage library displaying human single-chain variable fragments J. Biochem. 2010, 147(6), 809-817

２００９年

Hiroko Kawakami, Kohtaro Goto, Mamoru Mizuno "Multi-step synthesis of a protected monosaccharide unit by iterative reactions in microreactors and fluorous liquid-phase extractions" Chemistry Letters, 38(9), 906-907 (2009)

水野真盛 "フルオラス糖鎖合成の最近の展開" 複合糖質の化学と最新応用技術, pp19-27, シーエムシー出版 (2009)

Maria Carmelita Z. Kasuya, Mami Tojino, Shinya Nakano, Mamoru Mizuno and Kenichi Hatanaka "Effect of Fluorous Tags on Glycosylation of Saccharide Primers in Animal Cells" Bulletin of the Chemical Society of Japan, 82 (11), 1409-1415 (2009)