Skip to main content

Advertisement

SpringerLink
Log in

Simultaneous and sialic acid linkage-specific N- and O-linked glycan analysis by ester-to-amide derivatization

  • Research Article
  • Published:
Glycoconjugate Journal Aims and scope Submit manuscript

Abstract

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.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

Data availability

The data in this article will be available on reasonable request to the corresponding author.

References

  1. Surinova, S., Schiess, R., Hüttenhain, R., Cerciello, F., Wollscheid, B., Aebersold, R.: On the development of plasma protein biomarkers. J. Proteome Res. 10, 5–16 (2011). https://doi.org/10.1021/pr1008515

    Article  CAS  PubMed  Google Scholar 

  2. Varki, A., Cummings, R.D., Esko, J.D., Freeze, H.H., Stanley, P., Bertozzi, C.R., Hart, G.W., Etzler, M.E.: Essentials of Glycobiology, 2nd edn. Cold Spring Harbor Laboratory Press, NY (2009)

    Google Scholar 

  3. Plummer, T.H., Elder, J.H., Alexander, S., Phelan, A.W., Tarentino, A.L.: Demonstration of peptide: N-glycosidase F activity in endo-beta-N-acetylglucosaminidase F preparations. J. Biol. Chem. 259, 10700–10704 (1984)

    Article  CAS  PubMed  Google Scholar 

  4. Furukawa, J.I., Shinohara, Y., Kuramoto, H., Miura, Y., Shimaoka, H., Kurogochi, M., Nakano, M., Nishimura, S.: Comprehensive approach to structural and functional glycomics based on chemoselective glycoblotting and sequential tag conversion. Anal. Chem. 80, 1094–101 (2008). https://doi.org/10.1021/ac702124d

  5. Miura, Y., Shinohara, Y., Furukawa, J-i., Nagahori, N., Nishimura, S.: Rapid and simple solid-phase esterification of sialic acid residues for quantitative glycomics by mass spectrometry. Chemistry 13, 4797–804 (2007). https://doi.org/10.1002/chem.200601872

  6. Hanamatsu, H., Nishikaze, T., Miura, N., Piao, J., Okada, K., Sekiya, S., Iwamoto, S., Sakamoto, N., Tanaka, K., Furukawa, J.I.: Sialic Acid Linkage Specific Derivatization of Glycosphingolipid Glycans by Ring-Opening Aminolysis of Lactones. Anal. Chem. 90, 13193–13199 (2018). https://doi.org/10.1021/acs.analchem.8b02775

  7. Carlson, D.M.: Structures and immunochemical properties of oligosaccharides isolated from pig submaxillary mucins. J. Biol. Chem. 243, 616–626 (1968)

    Article  CAS  PubMed  Google Scholar 

  8. Huang, Y., Mechref, Y., Novotny, M.V.: Microscale nonreductive release of O-linked glycans for subsequent analysis through MALDI mass spectrometry and capillary electrophoresis. Anal. Chem. 73, 6063–6069 (2001). https://doi.org/10.1021/ac015534c

    Article  CAS  PubMed  Google Scholar 

  9. Miura, Y., Kato, K., Takegawa, Y., Kurogochi, M., Furukawa, J-i., Shinohara, Y., Nagahori, N., Amano, M., Hinou, H., Nishimura, S.I.: Glycoblotting-Assisted O-Glycomics: Ammonium Carbamate Allows for Highly Efficient O-Glycan Release from Glycoproteins. Anal. Chem. 82, 10021–10029 (2010). https://doi.org/10.1021/ac101599p

  10. Yu, G., Zhang, Y., Zhang, Z., Song, L., Wang, P., Chai, W.: Effect and limitation of excess ammonium on the release of O-glycans in reducing forms from glycoproteins under mild alkaline conditions for glycomic and functional analysis. Anal. Chem. 82, 9534–9542 (2010). https://doi.org/10.1021/ac102300r

    Article  CAS  PubMed  Google Scholar 

  11. Kameyama, A., Thet Tin, W.W., Toyoda, M., Sakaguchi, M.: A practical method of liberating O-linked glycans from glycoproteins using hydroxylamine and an organic superbase. Biochem Biophys Res Commun. 513, 186–192 (2019). https://doi.org/10.1016/j.bbrc.2019.03.144

    Article  CAS  PubMed  Google Scholar 

  12. Wheeler, S.F., Domann, P., Harvey, D.J.: Derivatization of sialic acids for stabilization in matrix-assisted laser desorption/ionization mass spectrometry and concomitant differentiation of α (2→ 3)-and α (2→ 6)-isomers. Rapid Communications in Mass Spectrometry: An International Journal Devoted to the Rapid Dissemination of Up-to-the-Minute Research in Mass Spectrometry. 23, 303–312 (2009)

    Article  CAS  Google Scholar 

  13. Alley, W.R. Jr., Novotny, M.V.: Glycomic analysis of sialic acid linkages in glycans derived from blood serum glycoproteins. J. Proteome Res. 9, 3062–3072 (2010)

  14. Reiding, K.R., Blank, D., Kuijper, D.M., Deelder, A.M., Wuhrer, M.: High-throughput profiling of protein N-glycosylation by MALDI-TOF-MS employing linkage-specific sialic acid esterification. Anal. Chem. 86, 5784–5793 (2014). https://doi.org/10.1021/ac500335t

    Article  CAS  PubMed  Google Scholar 

  15. de Haan, N., Reiding, K.R., Haberger, M., Reusch, D., Falck, D., Wuhrer, M.: Linkage-specific sialic acid derivatization for MALDI-TOF-MS profiling of IgG glycopeptides. Anal. Chem. 87, 8284–8291 (2015). https://doi.org/10.1021/acs.analchem.5b02426

    Article  CAS  PubMed  Google Scholar 

  16. Holst, S., Heijs, B., de Haan, N., van Zeijl, R.J., Briaire-de Bruijn, I.H., van Pelt, G.W., Mehta, A.S., Angel, P.M., Mesker, W.E., Tollenaar, R.A., Drake, R.R., Bovee, J.V., McDonnell, L.A., Wuhrer, M.: Linkage-Specific in Situ Sialic Acid Derivatization for N-Glycan Mass Spectrometry Imaging of Formalin-Fixed Paraffin-Embedded Tissues. Anal. Chem. 88, 5904–5913 (2016). https://doi.org/10.1021/acs.analchem.6b00819

    Article  CAS  PubMed  Google Scholar 

  17. Jin, W., Wang, C., Yang, M., Wei, M., Huang, L., Wang, Z.: Glycoqueuing: Isomer-Specific Quantification for Sialylation-Focused Glycomics. Anal. Chem. 91, 10492–10500 (2019). https://doi.org/10.1021/acs.analchem.9b01393

    Article  CAS  PubMed  Google Scholar 

  18. Peng, Y., Wang, L., Zhang, Y., Bao, H., Lu, H.: Stable Isotope Sequential Derivatization for Linkage-Specific Analysis of Sialylated N-Glycan Isomers by MS. Anal. Chem. 91, 15993–16001 (2019). https://doi.org/10.1021/acs.analchem.9b04727

    Article  CAS  PubMed  Google Scholar 

  19. Uematsu, R., Furukawa, J.I., Nakagawa, H., Shinohara, Y., Deguchi, K., Monde, K., Nishimura, S.: High throughput quantitative glycomics and glycoform-focused proteomics of murine dermis and epidermis. Mol. Cell Proteomics. 4, 1977–89 (2005). https://doi.org/10.1074/mcp.M500203-MCP200

  20. Fujitani, N., Takegawa, Y., Ishibashi, Y., Araki, K., Furukawa, J.I., Mitsutake, S., Igarashi, Y., Ito, M., Shinohara, Y.: Qualitative and quantitative cellular glycomics of glycosphingolipids based on rhodococcal endoglycosylceramidase-assisted glycan cleavage, glycoblotting-assisted sample preparation, and matrix-assisted laser desorption ionization tandem time-of-flight mass spectrometry analysis. J. Biol. Chem. 286, 41669–79 (2011). https://doi.org/10.1074/jbc.M111.301796

  21. Furukawa, J.I., Hanamatsu, H., Nishikaze, T., Manya, H., Miura, N., Yagi, H., Yokota, I., Akasaka-Manya, K., Endo, T., Kanagawa, M., Iwasaki, N., Tanaka, K.: Lactone-Driven Ester-to-Amide Derivatization for Sialic Acid Linkage-Specific Alkylamidation. Anal. Chem. 92, 14383–14392 (2020). https://doi.org/10.1021/acs.analchem.0c02209

  22. Hanamatsu, H., Nishikaze, T., Furukawa, J.I.: Comprehensive Glycan Analysis of Sphingolipids in Human Serum/Plasma. Methods Mol. Biol. 2613, 289–299 (2023). https://doi.org/10.1007/978-1-0716-2910-9_21

  23. Nishikaze, T., Tsumoto, H., Sekiya, S., Iwamoto, S., Miura, Y., Tanaka, K.: Differentiation of Sialyl Linkage Isomers by One-Pot Sialic Acid Derivatization for Mass Spectrometry-Based Glycan Profiling. Anal. Chem. 89, 2353–2360 (2017). https://doi.org/10.1021/acs.analchem.6b04150

    Article  CAS  PubMed  Google Scholar 

  24. Xu, L., Hanamatsu, H., Homan, K., Onodera, T., Miyazaki, T., Furukawa, J.I., Hontani, K., Tian, Y., Baba, R., Iwasaki, N.: Alterations of Glycosphingolipid Glycans and Chondrogenic Markers during Differentiation of Human Induced Pluripotent Stem Cells into Chondrocytes. Biomolecules 10 (2020). https://doi.org/10.3390/biom10121622

  25. Kita, Y., Miura, Y., Furukawa, J-i., Nakano, M., Shinohara, Y., Ohno, M., Takimoto, A., Nishimura, S.: Quantitative glycomics of human whole serum glycoproteins based on the standardized protocol for liberating N-glycans. Mol. Cellular Proteom. 6, 1437–1445 (2007). https://doi.org/10.1074/mcp.T600063-MCP200

  26. Yang, S., Wu, W.W., Shen, R., Sjogren, J., Parsons, L., Cipollo, J.F.: Optimization of O- GIG for O-Glycopeptide Characterization with Sialic Acid Linkage Determination. Anal. Chem. 92, 10946–10951 (2020). https://doi.org/10.1021/acs.analchem.0c01346

    Article  CAS  PubMed  Google Scholar 

  27. Lee, M.R., Shin, I.: Facile preparation of carbohydrate microarrays by site-specific, covalent immobilization of unmodified carbohydrates on hydrazide-coated glass slides. Org. Lett. 7, 4269–4272 (2005). https://doi.org/10.1021/ol051753z

    Article  CAS  PubMed  Google Scholar 

  28. Toegel, S., Bieder, D., André, S., Altmann, F., Walzer, S.M., Kaltner, H., Hofstaetter, J.G., Windhager, R., Gabius, H.J.: Glycophenotyping of osteoarthritic cartilage and chondrocytes by RT-qPCR, mass spectrometry, histochemistry with plant/human lectins and lectin localization with a glycoprotein. Arthritis Res. Ther. 15, R147 (2013). https://doi.org/10.1186/ar4330

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Funding

This work was supported by research funds from Sumitomo Electric Industries Ltd. (SEI), the Group CSR Foundation. This work was also supported in part by JSPS KAKENHI Grant Numbers 22H03502.

Author information

Authors and Affiliations

  1. Department of Orthopaedic Surgery, Faculty of Medicine and Graduate School of Medicine, Hokkaido University, Kita 15, Nishi 7, Kita-ku, Hokkaido, 060-8638, Sapporo, Japan

    Hisatoshi Hanamatsu, Kentaro Homan, Tomohiro Onodera, Norimasa Iwasaki & Jun-ichi Furukawa

  2. Department of Gastroenterology and Hepatology, Graduate School of Medicine, Hokkaido University, Sapporo, Hokkaido, 060-8638, Japan

    Hisatoshi Hanamatsu

  3. Sumitomo Bakelite Co., Ltd., 5-8, Tennoz Parkside Building, Higashi-Shinagawa 2-chome, Shinagawa-ku, 140-0002, Tokyo, Japan

    Yoshiaki Miura

  4. Solutions COE, Analytical & Measuring Instruments Division, Shimadzu Corporation, 604-8511, Kyoto, Japan

    Takashi Nishikaze & Yoshihiro Hayakawa

  5. Institute for Glyco-core Research (iGCORE), Nagoya University, 464-8601, Nagoya, Japan

    Ikuko Yokota & Jun-ichi Furukawa

Authors
  1. Hisatoshi Hanamatsu
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Yoshiaki Miura
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takashi Nishikaze
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ikuko Yokota
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Kentaro Homan
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Tomohiro Onodera
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Yoshihiro Hayakawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Norimasa Iwasaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Jun-ichi Furukawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization: Hisatoshi Hanamatsu, and Jun-ichi Furukawa; Methodology: Hisatoshi Hanamatsu, Yoshiaki Miura, Takashi Nishikaze, and Jun-ichi Furukawa; Formal analysis and investigation: Hisatoshi Hanamatsu, Ikuko Yokota, and Kentaro Homan; Writing -original draft preparation: Hisatoshi Hanamatsu, and Jun-ichi Furukawa; Writing review and editing: Hisatoshi Hanamatsu, Yoshiaki Miura, Takashi Nishikaze, Tomohiro Onodera, Yoshihiro Hayakawa, Norimasa Iwasaki, and Jun-ichi Furukawa; Supervision: Norimasa Iwasaki, and Jun-ichi Furukawa.

Corresponding authors

Correspondence to Hisatoshi Hanamatsu or Jun-ichi Furukawa.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Ethical approval

Acquisition and use of patient tissues were approved by the Institutional Review Board (IRB) of Hokkaido University (approval number: 014–0144), and informed consent was obtained in advance.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (PDF 712 KB)

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Hanamatsu, H., Miura, Y., Nishikaze, T. et al. Simultaneous and sialic acid linkage-specific N- and O-linked glycan analysis by ester-to-amide derivatization. Glycoconj J 40, 259–267 (2023). https://doi.org/10.1007/s10719-023-10109-8

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10719-023-10109-8

Keywords

Search

Navigation