De Novo Glycan Display on Cell Surfaces Using HaloTag: Visualizing the Effect of the Galectin Lattice on the Lateral Diffusion and Extracellular Vesicle Loading of Glycosylated Membrane ProteinsClick to copy article linkArticle link copied!
- Ayane MiuraAyane MiuraDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Ayane Miura
- Yoshiyuki Manabe*Yoshiyuki Manabe*Email: [email protected]. Phone: +81-6850-5391.Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanForefront Research Center, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Yoshiyuki Manabe
- Kenichi G. N. SuzukiKenichi G. N. SuzukiInstitute for Glyco-core Research (iGCORE), Gifu University, Gifu 501-1193, JapanNational Cancer Center Research Institute, Chuo-ku, Tokyo 104-0045, JapanMore by Kenichi G. N. Suzuki
- Hiroki ShomuraHiroki ShomuraDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Hiroki Shomura
- Soichiro OkamuraSoichiro OkamuraDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Soichiro Okamura
- Asuka ShirakawaAsuka ShirakawaDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Asuka Shirakawa
- Kumpei YanoKumpei YanoDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Kumpei Yano
- Shuto MiyakeShuto MiyakeDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Shuto Miyake
- Koki MayusumiKoki MayusumiDepartment of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanMore by Koki Mayusumi
- Chun-Cheng LinChun-Cheng LinDepartment of Chemistry, National Tsing-Hua University, Hsinchu 30013, TaiwanDepartment of Medicinal and Applied Chemistry, Kaohsiung Medical University, Kaohsiung 80708, TaiwanMore by Chun-Cheng Lin
- Kenta MorimotoKenta MorimotoDepartment of Biological Chemistry, Graduate School of Science, Osaka Metropolitan University, 1-1, Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, JapanMore by Kenta Morimoto
- Jojiro IshitobiJojiro IshitobiDepartment of Biological Chemistry, Graduate School of Science, Osaka Metropolitan University, 1-1, Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, JapanMore by Jojiro Ishitobi
- Ikuhiko NakaseIkuhiko NakaseDepartment of Biological Chemistry, Graduate School of Science, Osaka Metropolitan University, 1-1, Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, JapanDepartment of Biological Chemistry, School of Science, Osaka Metropolitan University, 1-1, Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, JapanMore by Ikuhiko Nakase
- Kenta AraiKenta AraiAdvanced ICT Research Institute, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe, Hyogo 651-2492, JapanMore by Kenta Arai
- Shouhei KobayashiShouhei KobayashiAdvanced ICT Research Institute, National Institute of Information and Communications Technology, 588-2 Iwaoka, Iwaoka-cho, Nishi-ku, Kobe, Hyogo 651-2492, JapanMore by Shouhei Kobayashi
- Ushio IshikawaUshio IshikawaDivision of Glycopathology, Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, 4-4-1, Komatsushima, Aoba Ward, Sendai, Miyagi 981-8558, JapanMore by Ushio Ishikawa
- Hirotaka KanohHirotaka KanohDivision of Glycopathology, Institute of Molecular Biomembrane and Glycobiology, Tohoku Medical and Pharmaceutical University, 4-4-1, Komatsushima, Aoba Ward, Sendai, Miyagi 981-8558, JapanMore by Hirotaka Kanoh
- Eiji MiyoshiEiji MiyoshiDepartment of Molecular Biochemistry and Clinical Investigation, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, JapanMore by Eiji Miyoshi
- Toshiyuki YamajiToshiyuki YamajiDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, Shinjuku-ku, Tokyo 162-8640, JapanMore by Toshiyuki Yamaji
- Kazuya Kabayama*Kazuya Kabayama*Email: [email protected]. Phone: +81-6850-5192.Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanForefront Research Center, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanInterdisciplinary Research Center for Radiation Sciences, Institute for Radiation Sciences, Osaka University, 2-4 Yamadaoka, Suita, Osaka 565-0871, JapanMore by Kazuya Kabayama
- Koichi Fukase*Koichi Fukase*Email: [email protected]. Phone: +81-6850-5391.Department of Chemistry, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanForefront Research Center, Graduate School of Science, Osaka University, 1-1 Machikaneyama, Toyonaka, Osaka 560-0043, JapanCenter for Advanced Modalities and DDS, Osaka University, 1-1 Yamadaoka, Suita, Osaka 565-0871, JapanMore by Koichi Fukase
Abstract
Glycans cover the cell surface to form the glycocalyx, which governs a myriad of biological phenomena. However, understanding and regulating glycan functions is extremely challenging due to the large number of heterogeneous glycans that engage in intricate interaction networks with diverse biomolecules. Glycocalyx-editing techniques offer potent tools to probe their functions. In this study, we devised a HaloTag-based technique for glycan manipulation, which enables the introduction of chemically synthesized glycans onto a specific protein (protein of interest, POI) and concurrently incorporates fluorescent units to attach homogeneous, well-defined glycans to the fluorescence-labeled POIs. Leveraging this HaloTag-based glycan-display system, we investigated the influence of the interactions between Gal-3 and various N-glycans on protein dynamics. Our analyses revealed that glycosylation modulates the lateral diffusion of the membrane proteins in a structure-dependent manner through interaction with Gal-3, particularly in the context of the Gal-3-induced formation of the glycan network (galectin lattice). Furthermore, N-glycan attachment was also revealed to have a significant impact on the extracellular vesicle-loading of membrane proteins. Notably, our POI-specific glycan introduction does not disrupt intact glycan structures, thereby enabling a functional analysis of glycans in the presence of native glycan networks. This approach complements conventional glycan-editing methods and provides a means for uncovering the molecular underpinnings of glycan functions on the cell surface.
Cited By
This article is cited by 1 publications.
- Pritam Ghosh. Deciphering the Cell Surface Sugar‐Coating via Biochemical Pathways. Chemistry – A European Journal 2024, 30
(64)
https://doi.org/10.1002/chem.202401983
- Koichiro M. Hirosawa, Yusuke Sato, Rinshi S. Kasai, Eriko Yamaguchi, Naoko Komura, Hiromune Ando, Ayuko Hoshino, Yasunari Yokota, Kenichi G. N. Suzuki. Uptake of small extracellular vesicles by recipient cells is facilitated by paracrine adhesion signaling. 2024https://doi.org/10.1101/2024.04.11.589012
Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.
Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.
The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.