Identification of Siglec Ligands Using a Proximity Labeling Method
| Autor: | Albert Ventura, Penk-Yeir Low, Chin-Ju Tang, Yu-Ju Chen, Chan-Yo Fan, Chun-Cheng Lin, Yi-Hsiu Chen, Takashi Angata, Lan-Yi Chang, Yi-Ju Chen |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
0301 basic medicine
Glycan Free Radicals Sialic Acid Binding Ig-like Lectin 2 Biotin Tyramine Biology Ligands Sialidase Proteomics Biochemistry Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Lectins Animals Humans Sialic Acid Binding Immunoglobulin-like Lectins B-Lymphocytes Staining and Labeling CD22 Pattern recognition receptor SIGLEC Lectin Hydrogen Peroxide General Chemistry respiratory system Sialic acid RAW 264.7 Cells 030104 developmental biology Immunoglobulin M chemistry Immune System 030220 oncology & carcinogenesis biology.protein Leukocyte Common Antigens |
| Zdroj: | Journal of Proteome Research. 16:3929-3941 |
| ISSN: | 1535-3907 1535-3893 |
| DOI: | 10.1021/acs.jproteome.7b00625 |
| Popis: | Siglecs are a family of receptor-type glycan recognition proteins (lectins) involved in self-nonself discrimination by the immune system. Identification of Siglec ligands is necessary to understand how Siglec-ligand interaction translates into biological outcomes. However, this is challenging because the interaction is weak. To facilitate identification of Siglec ligands, we adopted a proximity labeling method based on the tyramide radicalization principle. Cells that express Siglec ligands were labeled with Siglec-peroxidase complexes and incubated with biotin tyramide and hydrogen peroxide to generate short-lived tyramide radicals that covalently label the proteins near the Siglec-peroxidase complex. A proof-of-principle experiment using CD22 (Siglec-2) probe identified its known ligands on B cells, including CD22 itself, CD45, and IgM, among others, demonstrating the validity of this method. The specificity of labeling was confirmed by sialidase treatment of target cells and using glycan recognition-deficient mutant CD22 probes. Moreover, possible interactions between biotin-labeled proteins were revealed by literature-based protein-protein interaction network analysis, implying the presence of a molecular cluster comprising CD22 ligands. Further application of this method identified CD44 as a hitherto unknown Siglec-15 ligand on RAW264.7-derived osteoclasts. These results demonstrated the utility of proximity labeling for the identification of Siglec ligands, which may extend to other lectins. |
| Databáze: | OpenAIRE |
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