ACS Publications. Most Trusted. Most Cited. Most Read
My Activity

Figure 1Loading Img

GalNAc-Tyrosine Is a Ligand of Plant Lectins, Antibodies, and Human and Murine Macrophage Galactose-Type Lectins

View Author Information
Chemical Biology Laboratory, National Cancer Institute, National Institutes of Health, 376 Boyles St., Frederick, Maryland 21702, United States
*Tel.: (301) 846-5699. Fax: (301) 876-6033. E-mail: [email protected]
Cite this: ACS Chem. Biol. 2017, 12, 8, 2172–2182
Publication Date (Web):June 23, 2017
Copyright © 2017 American Chemical Society

    Article Views





    Other access options
    Supporting Info (2)»


    Abstract Image

    In 2011, a new type of protein O-glycosylation was discovered in which N-acetylgalactosamine is attached to the side chain of tyrosine (GalNAc-Tyr). While present on dozens of proteins, the biological roles of GalNAc-Tyr are unknown. To gain insight into this new type of modification, we synthesized a group of GalNAc-Tyr glycopeptides, constructed microarrays, and evaluated potential recognition of GalNAc-Tyr by a series of glycan-binding proteins. Through a series of >150 microarray experiments, we assessed binding properties of a variety of plant lectins, monoclonal antibodies, and endogenous lectins. VVL, HPA, and SBA were all found to bind tightly to GalNAc-Tyr, and several Tn binding antibodies and blood group A antibodies were found to cross-react with GalNAc-Tyr. Thus, detection of GalNAc-Tyr modified proteins is an important consideration when analyzing results from these reagents. Additionally, we evaluated potential recognition by two mammalian lectins, human (hMGL) and murine (mMGL-2) macrophage galactose type C-type lectins. Both hMGL and mMGL-2 bound tightly to GalNAc-Tyr determinants. The apparent Kd values (∼1–40 nM) were on par with some of the best known ligands for MGL, such as the Tn antigen. hMGL also bound the natural beta-amyloid peptide containing a GalNAc-Tyr epitope. STD NMR experiments provided structural insights into the molecular basis of recognition. Finally, GalNAc-Tyr was selectively captured by mMGL-2 positive dendritic cells. These results provide the first evidence that GalNAc-Tyr modified proteins and/or peptides may be ligands for hMGL and mMGL-2 and offer unique structures for the design of MGL targeting agents.

    Read this article

    To access this article, please review the available access options below.

    Get instant access

    Purchase Access

    Read this article for 48 hours. Check out below using your ACS ID or as a guest.


    Access through Your Institution

    You may have access to this article through your institution.

    Your institution does not have access to this content. You can change your affiliated institution below.

    Supporting Information

    Jump To

    The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acschembio.7b00471.

    • Experimental details and characterization data for all new compounds, ELISA data, representative array images, binding curves and apparent Kd values, flow cytometry data, and STD NMR data (PDF)

    • Tables of A167 focused array data (XLSX)

    Terms & Conditions

    Most electronic Supporting Information files are available without a subscription to ACS Web Editions. Such files may be downloaded by article for research use (if there is a public use license linked to the relevant article, that license may permit other uses). Permission may be obtained from ACS for other uses through requests via the RightsLink permission system:

    Cited By

    This article is cited by 22 publications.

    1. Zacharias Wijfjes, Floris J. van Dalen, Camille M. Le Gall, Martijn Verdoes. Controlling Antigen Fate in Therapeutic Cancer Vaccines by Targeting Dendritic Cell Receptors. Molecular Pharmaceutics 2023, 20 (10) , 4826-4847.
    2. Ignacio Sanz-Martinez, Sandra Pereira, Pedro Merino, Francisco Corzana, Ramon Hurtado-Guerrero. Molecular Recognition of GalNAc in Mucin-Type O-Glycosylation. Accounts of Chemical Research 2023, 56 (5) , 548-560.
    3. Li Xia, Tiffany R. Bellomo, Ruslan Gibadullin, Molly D. Congdon, Elijah F. Edmondson, Mi Li, Alexander Wlodawer, Crystal Li, J. Sebastian Temme, Pavan Patel, Donna Butcher, Jeffrey C. Gildersleeve. Development of a GalNAc-Tyrosine-Specific Monoclonal Antibody and Detection of Tyrosine O-GalNAcylation in Numerous Human Tissues and Cell Lines. Journal of the American Chemical Society 2022, 144 (36) , 16410-16422.
    4. Suraj Toraskar, Preeti Madhukar Chaudhary, Raghavendra Kikkeri. The Shape of Nanostructures Encodes Immunomodulation of Carbohydrate Antigen and Vaccine Development. ACS Chemical Biology 2022, 17 (5) , 1122-1130.
    5. Kevin R. Trabbic, Kristopher A. Kleski, Joseph J. Barchi, Jr.. Stable Gold-Nanoparticle-Based Vaccine for the Targeted Delivery of Tumor-Associated Glycopeptide Antigens. ACS Bio & Med Chem Au 2021, 1 (1) , 31-43.
    6. Molly D. Congdon, Jeffrey C. Gildersleeve. Enhanced Binding and Reduced Immunogenicity of Glycoconjugates Prepared via Solid-State Photoactivation of Aliphatic Diazirine Carbohydrates. Bioconjugate Chemistry 2021, 32 (1) , 133-142.
    7. Sarah V. Durbin, W. Shea Wright, Jeffrey C. Gildersleeve. Development of a Multiplex Glycan Microarray Assay and Comparative Analysis of Human Serum Anti-Glycan IgA, IgG, and IgM Repertoires. ACS Omega 2018, 3 (12) , 16882-16891.
    8. Gerard Artigas, João T. Monteiro, Hiroshi Hinou, Shin-Ichiro Nishimura, Bernd Lepenies, Fayna Garcia-Martin. Glycopeptides as Targets for Dendritic Cells: Exploring MUC1 Glycopeptides Binding Profile toward Macrophage Galactose-Type Lectin (MGL) Orthologs. Journal of Medicinal Chemistry 2017, 60 (21) , 9012-9021.
    9. Sandra Behren, Manuel Schorlemer, Gudula Schmidt, Klaus Aktories, Ulrika Westerlind. Antibodies Directed Against GalNAc‐ and GlcNAc‐ O‐ Tyrosine Posttranslational Modifications – a New Tool for Glycoproteomic Detection. Chemistry – A European Journal 2023, 29 (29)
    10. Yujun Kim, Ji Young Hyun, Injae Shin. Glycan microarrays from construction to applications. Chemical Society Reviews 2022, 51 (19) , 8276-8299.
    11. Shuhui Chen, Rui Qin, Lara K. Mahal. Sweet systems: technologies for glycomic analysis and their integration into systems biology. Critical Reviews in Biochemistry and Molecular Biology 2021, 56 (3) , 301-320.
    12. Pablo Valverde, J. Daniel Martínez, F. Javier Cañada, Ana Ardá, Jesús Jiménez‐Barbero. Molecular Recognition in C‐Type Lectins: The Cases of DC‐SIGN, Langerin, MGL, and L‐Sectin. ChemBioChem 2020, 21 (21) , 2999-3025.
    13. J. Kenneth Hoober. ASGR1 and Its Enigmatic Relative, CLEC10A. International Journal of Molecular Sciences 2020, 21 (14) , 4818.
    14. Agnieszka J. Pietrzyk-Brzezinska, Anna Bujacz. H-type lectins – Structural characteristics and their applications in diagnostics, analytics and drug delivery. International Journal of Biological Macromolecules 2020, 152 , 735-747.
    15. Amedeo Amedei, Fatemeh Asadzadeh, Francesco Papi, Maria Giuliana Vannucchi, Veronica Ferrucci, Iris A. Bermejo, Marco Fragai, Carolina Vieira De Almeida, Linda Cerofolini, Stefano Giuntini, Mauro Bombaci, Elisa Pesce, Elena Niccolai, Francesca Natali, Eleonora Guarini, Frank Gabel, Chiara Traini, Stefano Catarinicchia, Federica Ricci, Lorenzo Orzalesi, Francesco Berti, Francisco Corzana, Massimo Zollo, Renata Grifantini, Cristina Nativi. A Structurally Simple Vaccine Candidate Reduces Progression and Dissemination of Triple-Negative Breast Cancer. iScience 2020, 23 (6) , 101250.
    16. Martina Pirro, Yoann Rombouts, Alexandre Stella, Olivier Neyrolles, Odile Burlet-Schiltz, Sandra J. van Vliet, Arnoud H. de Ru, Yassene Mohammed, Manfred Wuhrer, Peter A. van Veelen, Paul J. Hensbergen. Characterization of Macrophage Galactose-type Lectin (MGL) ligands in colorectal cancer cell lines. Biochimica et Biophysica Acta (BBA) - General Subjects 2020, 1864 (4) , 129513.
    17. Kristopher A. Kleski, Kevin R. Trabbic, Mengchao Shi, Jean-Paul Bourgault, Peter R. Andreana. Enhanced Immune Response Against the Thomsen-Friedenreich Tumor Antigen Using a Bivalent Entirely Carbohydrate Conjugate. Molecules 2020, 25 (6) , 1319.
    18. Richard D. Cummings. “Stuck on sugars – how carbohydrates regulate cell adhesion, recognition, and signaling”. Glycoconjugate Journal 2019, 36 (4) , 241-257.
    19. Zhonghua Li, Tatiana A. Chernova, Tongzhong Ju. Novel Technologies for Quantitative O -Glycomics and Amplification/Preparation of Cellular O -Glycans. 2019, 370-392.
    20. Filipa Marcelo, Nitin Supekar, Francisco Corzana, Joost C. van der Horst, Ilona M. Vuist, David Live, Geert-Jan P.H. Boons, David F. Smith, Sandra J. van Vliet. Identification of a secondary binding site in human macrophage galactose-type lectin by microarray studies: Implications for the molecular recognition of its ligands. Journal of Biological Chemistry 2019, 294 (4) , 1300-1311.
    21. David Montoir, Mehdi Amoura, Zine El Abidine Ababsa, T. M. Vishwanatha, Expédite Yen-Pon, Vincent Robert, Massimiliano Beltramo, Véronique Piller, Mouad Alami, Vincent Aucagne, Samir Messaoudi. Synthesis of aryl-thioglycopeptides through chemoselective Pd-mediated conjugation. Chemical Science 2018, 9 (46) , 8753-8759.
    22. Dorota Hoja-Łukowicz, Sabina Szwed, Piotr Laidler, Anna Lityńska. Proteomic analysis of Tn-bearing glycoproteins from different stages of melanoma cells reveals new biomarkers. Biochimie 2018, 151 , 14-26.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    Pair your accounts.

    Export articles to Mendeley

    Get article recommendations from ACS based on references in your Mendeley library.

    You’ve supercharged your research process with ACS and Mendeley!

    STEP 1:
    Click to create an ACS ID

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Please note: If you switch to a different device, you may be asked to login again with only your ACS ID.

    Your Mendeley pairing has expired. Please reconnect