Chemokine binding to PSGL-1 is controlled by O-glycosylation and tyrosine sulfation
Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
Protein glycosylation influences cellular recognition and regulates protein interactions, but how glycosylation functions alongside other common posttranslational modifications (PTMs), like tyrosine sulfation (sTyr), is unclear. We produced a library of 53 chemoenzymatically synthesized glycosulfopeptides representing N-terminal domains of human and murine P-selectin glycoprotein ligand-1 (PSGL-1), varying in sTyr and O-glycosylation (structure and site). Using these, we identified key roles of PSGL-1 O-glycosylation and sTyr in controlling interactions with specific chemokines. Results demonstrate that sTyr positively affects CCL19 and CCL21 binding to PSGL-1 N terminus, whereas O-glycan branching and sialylation reduced binding. For murine PSGL-1, interference between PTMs is greater, attributed to proximity between the two PTMs. Using fluorescence polarization, we found sTyr is a positive determinant for some chemokines. We showed that synthetic sulfopeptides are potent in decreasing chemotaxis of human dendritic cells toward CCL19 and CCL21. Our results provide new research avenues into the interplay of PTMs regulating leukocyte/chemokine interactions.
| Originalsprog | Engelsk |
|---|---|
| Tidsskrift | Cell Chemical Biology |
| Vol/bind | 30 |
| Udgave nummer | 8 |
| Sider (fra-til) | 893-905.e7 |
| ISSN | 2451-9456 |
| DOI | |
| Status | Udgivet - 2023 |
Bibliografisk note
Funding Information:
This work was supported by the independent research fund Denmark ( 7025-00083B ) to C.K.G. and the Lundbeck Foundation ( R322-2019-2171 ) to C.K.G., and by NIH grants P41GM103694 and R24GM137763 to R.D.C. The authors would like to thank Dr. Kelley Moremen (University of Georgia) and the Repository of Glyco-enzyme Expression Constructs ( http://glycoenzymes.ccrc.uga.edu/ ) (the National Institutes of Health Grant P41GM103390 and P01GM107012 ) for the glycosyltransferase constructs.
Funding Information:
This work was supported by the independent research fund Denmark (7025-00083B) to C.K.G. and the Lundbeck Foundation (R322-2019-2171) to C.K.G. and by NIH grants P41GM103694 and R24GM137763 to R.D.C. The authors would like to thank Dr. Kelley Moremen (University of Georgia) and the Repository of Glyco-enzyme Expression Constructs (http://glycoenzymes.ccrc.uga.edu/) (the National Institutes of Health Grant P41GM103390 and P01GM107012) for the glycosyltransferase constructs. C.K.G. and A.Y.M. devised the synthesis methodology, performed the synthesis, MALDI/HPLC characterization and purification. A.Y.M. performed the printing of the array. C.K.G. performed enzymatic extension of peptides and the fluorescence polarization assay. A.Y.M. C.K.G. A.M.M. and K.J.B. performed the microarray experiments. M.S.H. and S.S.P. developed and synthesized the sulfonate containing peptides and microarray. K.S. performed the MS/MS experiments and analysis. G.M.H. and C.K.G. carried out the chemotaxis experiments. R.D.C. supervised the experiments and guided the project. J.H.M. E.L.C. and M.M.R. provided input and helped with interpreting results and planning experiments. A.Y.M. C.K.G. J.H.M. and R.D.C. wrote the article, and all authors edited and approved the final manuscript. The authors declare no competing interests. We support inclusive, diverse, and equitable conduct of research.
Publisher Copyright:
© 2023 Elsevier Ltd
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