The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis
Proinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential...
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MDPI AG
2022-06-01
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| Series: | International Journal of Molecular Sciences |
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| author | Alexandra Pum Maria Ennemoser Tanja Gerlza Andreas J. Kungl |
| author_facet | Alexandra Pum Maria Ennemoser Tanja Gerlza Andreas J. Kungl |
| author_sort | Alexandra Pum |
| collection | DOAJ |
| description | Proinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential co-receptors in the process of CCL26-induced eosinophil chemotaxis. For this purpose, we have first identified the GAG-binding site of CCL26 by a site-directed mutagenesis approach in the form of an alanine screening. A panel of GAG-binding-deficient mutants has been designed, generated, and analyzed with respect to their binding affinities to heparan sulphate (HS) by isothermal fluorescence titration studies. This showed that basic amino acids in the α-helical part of CCL26 are strongly involved in GAG-binding. In chemotaxis experiments, we found that decreased GAG-binding affinity correlated with decreased chemotactic activity, which indicates an involvement of GAGs in eosinophil migration. This was further proven by the negative impact of heparinase III treatment and, independently, by the incubation of eosinophils with an anti heparan sulfate antibody. We finally investigated eosinophils’ proteoglycan (PG) expression patterns by real-time PCR, which revealed the highest expression level for serglycin. Including an anti-serglycin antibody in CCL26-induced eosinophil migration experiments reduced the chemotaxis of these immune cells, thereby proving the dependence of eosinophil mobilization on the proteoglycan serglycin. |
| first_indexed | 2024-03-09T23:34:58Z |
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| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T23:34:58Z |
| publishDate | 2022-06-01 |
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| series | International Journal of Molecular Sciences |
| spelling | doaj.art-46a521b992874fa5842058e4c4c715862023-11-23T17:02:08ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-06-012312651910.3390/ijms23126519The Role of Heparan Sulfate in CCL26-Induced Eosinophil ChemotaxisAlexandra Pum0Maria Ennemoser1Tanja Gerlza2Andreas J. Kungl3Institute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, Karl-Franzens-University Graz, Schubertstrasse 1, 8010 Graz, AustriaInstitute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, Karl-Franzens-University Graz, Schubertstrasse 1, 8010 Graz, AustriaInstitute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, Karl-Franzens-University Graz, Schubertstrasse 1, 8010 Graz, AustriaInstitute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, Karl-Franzens-University Graz, Schubertstrasse 1, 8010 Graz, AustriaProinflammatory chemokine ligand 26 (CCL26, eotaxin-3) mediates transendothelial cell migration of eosinophils by binding and activating the G-protein-coupled (GPC) chemokine receptor 3 on the surface of eosinophilic cells. Here we have investigated the role of glycosaminoglycans (GAGs) as potential co-receptors in the process of CCL26-induced eosinophil chemotaxis. For this purpose, we have first identified the GAG-binding site of CCL26 by a site-directed mutagenesis approach in the form of an alanine screening. A panel of GAG-binding-deficient mutants has been designed, generated, and analyzed with respect to their binding affinities to heparan sulphate (HS) by isothermal fluorescence titration studies. This showed that basic amino acids in the α-helical part of CCL26 are strongly involved in GAG-binding. In chemotaxis experiments, we found that decreased GAG-binding affinity correlated with decreased chemotactic activity, which indicates an involvement of GAGs in eosinophil migration. This was further proven by the negative impact of heparinase III treatment and, independently, by the incubation of eosinophils with an anti heparan sulfate antibody. We finally investigated eosinophils’ proteoglycan (PG) expression patterns by real-time PCR, which revealed the highest expression level for serglycin. Including an anti-serglycin antibody in CCL26-induced eosinophil migration experiments reduced the chemotaxis of these immune cells, thereby proving the dependence of eosinophil mobilization on the proteoglycan serglycin.https://www.mdpi.com/1422-0067/23/12/6519CCL26eotaxin-3glycosaminoglycansGAGheparan sulfateheparinase C |
| spellingShingle | Alexandra Pum Maria Ennemoser Tanja Gerlza Andreas J. Kungl The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis International Journal of Molecular Sciences CCL26 eotaxin-3 glycosaminoglycans GAG heparan sulfate heparinase C |
| title | The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis |
| title_full | The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis |
| title_fullStr | The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis |
| title_full_unstemmed | The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis |
| title_short | The Role of Heparan Sulfate in CCL26-Induced Eosinophil Chemotaxis |
| title_sort | role of heparan sulfate in ccl26 induced eosinophil chemotaxis |
| topic | CCL26 eotaxin-3 glycosaminoglycans GAG heparan sulfate heparinase C |
| url | https://www.mdpi.com/1422-0067/23/12/6519 |
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