Abstract
Siglecs are known to be bound and regulated by membrane molecules that display specific sialic acid-containing ligands and are present on the same cell (cis-ligands). Because of the low-affinity binding of Siglecs to the glycan ligands, conventional methods such as immunoprecipitation are not suitable for identification of Siglec cis-ligands. Here we describe efficient and specific labeling of cis-ligands of CD22 (also known as Siglec-2) on B lymphocytes by proximity labeling using tyramide. This method may also be applicable to labeling of cis-ligands of other Siglecs.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Macauley MS, Crocker PR, Paulson JC (2014) Siglec-mediated regulation of immune cell function in disease. Nat Rev Immunol 14:653–666
Bobrow MN, Harris TD, Shaughnessy KJ et al (1989) Catalyzed reporter deposition, a novel method of signal amplification. Application to immunoassays. J Immunol Methods 125:279–285
Adams JC (1992) Biotin amplification of biotin and horseradish peroxidase signals in histochemical stains. J Histochem Cytochem 40:1457–1463
Kerstens HM, Poddighe PJ, Hanselaar AG (1995) A novel in situ hybridization signal amplification method based on the deposition of biotinylated tyramine. J Histochem Cytochem 43:347–352
Mayer G, Bendayan M (1997) Biotinyl-tyramide: a novel approach for electron microscopic immunocytochemistry. J Histochem Cytochem 45:1449–1454
Raap AK, van de Corput MP, Vervenne RA et al (1995) Ultra-sensitive FISH using peroxidase-mediated deposition of biotin- or fluorochrome tyramides. Hum Mol Genet 4:529–534
Schofer C, Weipoltshammer K, Almeder M et al (1997) Signal amplification at the ultrastructural level using biotinylated tyramides and immunogold detection. Histochem Cell Biol 108:313–319
Bendayan M (2001) Tech.Sight. Worth its weight in gold. Science 291:1363–1365
Li XW, Rees JS, Xue P et al (2014) New insights into the DT40 B cell receptor cluster using a proteomic proximity labeling assay. J Biol Chem 289:14434–14447
Rees JS, Li XW, Perrett S et al (2015) Protein neighbors and proximity proteomics. Mol Cell Proteomics 14:2848–2856
Chang L, Chen YJ, Fan CY et al (2017) Identification of Siglec ligands using a proximity labeling method. J Proteome Res 16:3929–3941
Alborzian Deh Sheikh A, Akatsu C, Imamura A et al (2018) Proximity labeling of cis-ligands of CD22/Siglec-2 reveals stepwise alpha2,6 sialic acid-dependent and -independent interactions. Biochem Biophys Res Commun 495:854–859
Hennet T, Chui D, Paulson JC et al (1998) Immune regulation by the ST6Gal sialyltransferase. Proc Natl Acad Sci U S A 95:4504–4509
Stoddart A, Ray RJ, Paige CJ (1997) Analysis of murine CD22 during B cell development: CD22 is expressed on B cell progenitors prior to IgM. Int Immunol 9:1571–1579
Abdu-Allah HH, Watanabe K, Completo GC et al (2011) CD22-antagonists with nanomolar potency: the synergistic effect of hydrophobic groups at C-2 and C-9 of sialic acid scaffold. Bioorg Med Chem 19:1966–1971
Jiang S, Kotani N, Ohnishi T et al (2012) A proteomics approach to the cell-surface interactome using the enzyme-mediated activation of radical sources reaction. Proteomics 12:54–62
Nomura T, Han H, Howard MC et al (1996) Antigen receptor-mediated B cell death is blocked by signaling via CD72 or treatment with dextran sulfate and is defective in autoimmunity-prone mice. Int Immunol 8:867–875
Acknowledgments
This work was funded by JPSP Grant-in-Aid for Scientific Research 26293062 (T.T.) and the Joint Usage/Research Program of Medical Research Institute, Tokyo Medical and Dental University (T.T.).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2020 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Alborzian Deh Sheikh, A., Akatsu, C., Tsubata, T. (2020). Identification of Siglec Cis-Ligands by Proximity Labeling. In: Hirabayashi, J. (eds) Lectin Purification and Analysis. Methods in Molecular Biology, vol 2132. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0430-4_8
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0430-4_8
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-0429-8
Online ISBN: 978-1-0716-0430-4
eBook Packages: Springer Protocols