Updated on 2023/10/06

写真a

 
FURUKAWA Junichi
 
Organization
Institute for Glyco-core Research Designated professor
Title
Designated professor
 

Papers 11

  1. Focal-adhesion kinase regulates the sialylation of N-glycans via the PI4KIIα-PI4P pathway. Reviewed International journal

    Yuhan Sun, Tomoya Isaji, Yoshiyuki Oyama, Xing Xu, Jianwei Liu, Hisatoshi Hanamatsu, Ikuko Yokota, Nobuaki Miura, Jun-Ichi Furukawa, Tomohiko Fukuda, Jianguo Gu

    The Journal of biological chemistry     page: 105051 - 105051   2023.7

     More details

    Language:English   Publishing type:Research paper (scientific journal)  

    Sialylation is a terminal glycosylated modification of glycoproteins that regulates critical biological events such as cell adhesion and immune response. Our previous study showed that integrin α3β1 plays a crucial role in regulating the sialylation of N-glycans. However, the underlying mechanism for the regulation remains unclear. This study investigated how sialylation is affected by focal adhesion kinase (FAK), which is a critical downstream signal molecule of integrin β1. We established a stable focal adhesion kinase (FAK) knockout (KO) cell line using the CRISPR/Cas9 system in HeLa cells. The results obtained from lectin blot, flow cytometric analysis, and mass spectrometry (MS) showed that the sialylation levels were significantly decreased in the KO cells compared with that in wild-type (WT) cells. Moreover, phosphatidylinositol 4-phosphate (PI4P) expression levels were also reduced in the KO cells due to a decrease in the stability of phosphatidylinositol 4-kinase-IIα (PI4KIIα). Notably, the decreased levels of sialylation, PI4P, and the complex formation between GOLPH3 and ST3GAL4 or ST6GAL1, which are the main sialyltransferases for modification of N-glycans, were significantly restored by the re-expression of FAK. Furthermore, the decreased sialylation and phosphorylation of Akt and cell migration caused by FAK deficiency all were restored by overexpressing PI4KIIα, which suggests that PI4KIIα is one of the downstream molecules of FAK. These findings indicate that FAK regulates sialylation via the PI4P synthesis pathway and a novel mechanism is suggested for the integrin-FAK-PI4KIIα-GOLPH3-ST axis modulation of sialylation in N-glycans.

    DOI: 10.1016/j.jbc.2023.105051

    PubMed

  2. Simultaneous and sialic acid linkage-specific N- and O-linked glycan analysis by ester-to-amide derivatization. Invited Reviewed International journal

    Hisatoshi Hanamatsu, Yoshiaki Miura, Takashi Nishikaze, Ikuko Yokota, Kentaro Homan, Tomohiro Onodera, Yoshihiro Hayakawa, Norimasa Iwasaki, Jun-Ichi Furukawa

    Glycoconjugate journal   Vol. 40 ( 2 ) page: 259 - 267   2023.4

     More details

    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Characterization of O-glycans linked to serine or threonine residues in glycoproteins has mostly been achieved using chemical reaction approaches because there are no known O-glycan-specific endoglycosidases. Most O-glycans are modified with sialic acid residues at the non-reducing termini through various linkages. In this study, we developed a novel approach for sialic acid linkage-specific O-linked glycan analysis through lactone-driven ester-to-amide derivatization combined with non-reductive β-elimination in the presence of hydroxylamine. O-glycans released by non-reductive β-elimination were efficiently purified using glycoblotting via chemoselective ligation between carbohydrates and a hydrazide-functionalized polymer, followed by modification of methyl or ethyl ester groups of sialic acid residues on solid-phase. In-solution lactone-driven ester-to-amide derivatization of ethyl-esterified O-glycans was performed, and the resulting sialylated glycan isomers were discriminated by mass spectrometry. In combination with PNGase F digestion, we carried out simultaneous, quantitative, and sialic acid linkage-specific N- and O-linked glycan analyses of a model glycoprotein and human cartilage tissue. This novel glycomic approach will facilitate detailed characterization of biologically relevant sialylated N- and O-glycans on glycoproteins.

    DOI: 10.1007/s10719-023-10109-8

    PubMed

  3. Exposure to brefeldin A induces unusual expression of hybrid- and complex-type free N-glycans in HepG2 cells Reviewed

    Kanako Sugiura, Yuho Kawai, Arisa Yamamoto, Hiroki Yoshioka, Yuika Kiyohara, Ayaka Iida, Yurika Ozawa, Mai Nishikawa, Nobuaki Miura, Hisatoshi Hanamatsu, Jun-ichi Furukawa, Yasuro Shinohara

    Biochimica et Biophysica Acta (BBA) - General Subjects   Vol. 1867 ( 5 ) page: 130331 - 130331   2023.2

     More details

    Language:English   Publishing type:Research paper (scientific journal)   Publisher:Elsevier BV  

    DOI: 10.1016/j.bbagen.2023.130331

    PubMed

  4. Comprehensive Glycan Analysis of Sphingolipids in Human Serum/Plasma. International journal

    Hisatoshi Hanamatsu, Takashi Nishikaze, Jun-Ichi Furukawa

    Methods in molecular biology (Clifton, N.J.)   Vol. 2613   page: 289 - 299   2023

     More details

    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Glycosphingolipids (GSLs) are glycolipids with ceramide and carbohydrate head groups that play an important role in numerous biological processes. Previously, we performed GSL-glycan analysis of various cell lines and virus-infected cells using a glycoblotting approach. Recently, we developed several methods for sialic acid linkage-specific chemical modification to distinguish sialylated glycan isomers by mass spectrometry. In this chapter, we describe a method for analyzing GSL-glycans in human serum/plasma using glycoblotting combined with aminolysis-SALSA (sialic acid linkage-specific alkylamidation) and lactone-driven ester-to-amide derivatization (LEAD)-SALSA for comprehensive and detailed structural glycomics.

    DOI: 10.1007/978-1-0716-2910-9_21

    PubMed

  5. Simultaneous determination of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan glycosaminoglycan disaccharides by high-performance liquid chromatography using a reverse-phase column with adamantyl groups. Reviewed International journal

    Hisatoshi Hanamatsu, Satoshi Makino, Masatsugu Ohara, Goki Suda, Ikuko Yokota, Shoko Nishihara, Naoya Sakamoto, Jun-Ichi Furukawa

    Journal of chromatography. A   Vol. 1689   page: 463748 - 463748   2022.12

     More details

    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    Glycosaminoglycans (GAGs), which are one of the major components of proteoglycans, play a pivotal role in physiological processes such as signal transduction, cell adhesion, growth, and differentiation. Characterization of GAGs is challenging due to the tremendous structural diversity of heteropolysaccharides with numerous sulfate or carboxyl groups. In this present study, we examined the analysis of 2-aminobenzamide (2-AB) labeled GAG disaccharides by high-performance liquid chromatography (HPLC) using a reverse-phase (RP)-column with adamantyl groups. Under the analytical conditions, 17 types of 2-AB labeled GAG disaccharides derived from heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan were sequentially separated in a single analysis. The analysis time was fast with high retention time reproducibility. Moreover, the RP-HPLC column with adamantyl groups allowed the quantification of GAGs in various biological samples, such as serum, cultured cells, and culture medium.

    DOI: 10.1016/j.chroma.2022.463748

    PubMed

  6. Establishment of the removal method of undifferentiated induced pluripotent stem cells coexisting with chondrocytes using R-17F antibody Reviewed

    Takuji Miyazaki, Hisatoshi Hanamatsu, Tomohiro Onodera, Jun-Ichi Furukawa, Liang Xu, Kentaro Homan, Rikiya Baba, Toshisuke Kawasaki, Norimasa Iwasaki

    Regenerative Medicine   Vol. 17 ( 11 ) page: 793 - 803   2022.11

     More details

    Authorship:Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: DOI: 10.2217/rme-2022-0010

  7. Toolbox Accelerating Glycomics (TAG): Improving Large-Scale Serum Glycomics and Refinement to Identify SALSA-Modified and Rare Glycans Invited Reviewed

    Nobuaki Miura 1, Hisatoshi Hanamatsu, Ikuko Yokota, Keiko Akasaka-Manya, Hiroshi Manya, Tamao Endo, Yasuro Shinohara and Jun-ichi Furukawa

    Int. J. Mol. Sci.   Vol. 23 ( 21 ) page: 13097   2022.10

     More details

    Authorship:Last author, Corresponding author   Language:English   Publishing type:Research paper (scientific journal)  

    DOI: https://doi.org/10.3390/ijms232113097

  8. Comprehensive Cellular Glycan Profiling of Glycoproteins and Glycosphingolipids by Glycoblotting and BEP Methods Invited

    Hisatoshi Hanamatsu, Jun-Ichi Furukawa

    Methods in Molecular Biology     page: 1 - 18   2022.9

     More details

    Authorship:Last author, Corresponding author   Language:English   Publishing type:Part of collection (book)  

    DOI: DOI: 10.1007/978-1-0716-2635-1_1

  9. Glycosphingolipid GM3 prevents albuminuria and podocytopathy induced by anti-nephrin antibody

    Kawashima Nagako, Naito Shokichi, Hanamatsu Hisatoshi, Nagane Masaki, Takeuchi Yasuo, Furukawa Jun-ichi, Iwasaki Norimasa, Yamashita Tadashi, Nakayama Ken-ichi

    SCIENTIFIC REPORTS   Vol. 12 ( 1 )   2022.9

     More details

    Language:Japanese   Publisher:Scientific Reports  

    Podocytopathy, which is characterized by injury to podocytes, frequently causes proteinuria or nephrotic syndrome. There is currently a paucity of effective therapeutic drugs to treat proteinuric kidney disease. Recent research suggests the possibility that glycosphingolipid GM3 maintains podocyte function by acting on various molecules including nephrin, but its mechanism of action remains unknown. Here, various analyses were performed to examine the potential relationship between GM3 and nephrin, and the function of GM3 in podocytes using podocytopathy mice, GM3 synthase gene knockout mice, and nephrin injury cells. Reduced amounts of GM3 and nephrin were observed in podocytopathy mice. Intriguingly, this reduction of GM3 and nephrin, as well as albuminuria, were inhibited by administration of valproic acid. However, when the same experiment was performed using GM3 synthase gene knockout mice, valproic acid administration did not inhibit albuminuria. Equivalent results were obtained in model cells. These findings indicate that GM3 acts with nephrin in a collaborative manner in the cell membrane. Taken together, elevated levels of GM3 stabilize nephrin, which is a key molecule of the slit diaphragm, by enhancing the environment of the cell membrane and preventing albuminuria. This study provides novel insight into new drug discovery, which may offer a new therapy for kidney disease with albuminuria.

    DOI: 10.1038/s41598-022-20265-w

    Web of Science

    Scopus

  10. Evaluation of the context of downstream N- and free N-glycomic alterations induced by swainsonine in HepG2 cells

    Morikawa Chie, Sugiura Kanako, Kondo Keina, Yamamoto Yurie, Kojima Yuma, Ozawa Yurika, Yoshioka Hiroki, Miura Nobuaki, Piao Jinhua, Okada Kazue, Hanamatsu Hisatoshi, Tsuda Masumi, Tanaka Shinya, Furukawa Jun-ichi, Shinohara Yasuro

    BIOCHIMICA ET BIOPHYSICA ACTA-GENERAL SUBJECTS   Vol. 1866 ( 9 )   2022.9

     More details

    Language:Japanese   Publisher:Biochimica et Biophysica Acta - General Subjects  

    Swainsonine (SWA), a potent inhibitor of class II α-mannosidases, is present in a number of plant species worldwide and causes severe toxicosis in livestock grazing these plants. The mechanisms underlying SWA-induced animal poisoning are not fully understood. In this study, we analyzed the alterations that occur in N- and free N-glycomic upon addition of SWA to HepG2 cells to understand better SWA-induced glycomic alterations. After SWA addition, we observed the appearance of SWA-specific glycomic alterations, such as unique fucosylated hybrid-type and fucosylated M5 (M5F) N-glycans, and a remarkable increase in all classes of Gn1 FNGs. Further analysis of the context of these glycomic alterations showed that (fucosylated) hybrid type N-glycans were not the precursors of these Gn1 FNGs and vice versa. Time course analysis revealed the dynamic nature of glycomic alterations upon exposure of SWA and suggested that accumulation of free N-glycans occurred earlier than that of hybrid-type N-glycans. Hybrid-type N-glycans, of which most were uniquely core fucosylated, tended to increase slowly over time, as was observed for M5F N-glycans. Inhibition of swainsonine-induced unique fucosylation of hybrid N-glycans and M5 by coaddition of 2-fluorofucose caused significant increases in paucimannose- and fucosylated paucimannose-type N-glycans, as well as paucimannose-type free N-glycans. The results not only revealed the gross glycomic alterations in HepG2 cells induced by swainsonine, but also provide information on the global interrelationships between glycomic alterations.

    DOI: 10.1016/j.bbagen.2022.130168

    Web of Science

    Scopus

  11. Comprehensive and Comparative Structural Glycome Analysis in Mouse Epiblast-like Cells Invited

    Federico Pecori, Hisatoshi Hanamatsu, Jun-ichi Furukawa, Shoko Nishihara

    Methods in Molecular Biology     page: 179 - 193   2022.4

     More details

    Authorship:Corresponding author   Language:English   Publishing type:Part of collection (book)  

    DOI: DOI: 10.1007/978-1-0716-2281-0_13

▼display all

KAKENHI (Grants-in-Aid for Scientific Research) 1

  1. 包括的組織グライコームによる糖鎖分子ネットワークの構築と老化の客観的評価

    Grant number:22H03502  2022.4 - 2026.3

    科学研究費助成事業  基盤研究(B)

    古川 潤一, 萬谷 博, 三浦 信明

      More details

    Authorship:Principal investigator 

    Grant amount:\17420000 ( Direct Cost: \13400000 、 Indirect Cost:\4020000 )

    申請者はこれまでに特定のサブグライコームだけではなく、種々のサブグライコームを解析する技術の開発によって、細胞や血清の包括的なグライコームを利用した網羅的なバイオマーカ探索等を行ってきた。本申請課題では、申請者が有する独自技術を基盤として組織や体液の包括的なグライコーム(全糖鎖)解析法を確立し、組織横断的な総合グライコームを明らかにする。さらに老化に伴うグライコーム変化と糖鎖合成・代謝に関わる遺伝子発現変動を統合した糖鎖分子ネットワーク変化を指標とした老化の客観的評価法について精査する。