Glycosylation Regulates N-Terminal Proteolysis and Activity of the Chemokine CCL14

Wang, Siyao; Foster, Simon R.; Sanchez, Julie; Corcilius, Leo; Larance, Mark; Canals, Meritxell; Stone, Martin J.; Payne, Richard J.

Abstract

Chemokines are secreted proteins that regulate leukocyte migration during inflammatory responses by signaling through chemokine receptors. Full length CC chemokine ligand 14, CCL14(1–74), is a weak agonist for the chemokine receptor CCR1, but its activity is substantially enhanced upon proteolytic cleavage to CCL14(9–74). CCL14 is O-glycosylated at Ser7, adjacent to the site of proteolytic activation. To determine whether glycosylation regulates the activity of CCL14, we used native chemical ligation to prepare four homogeneously glycosylated variants of CCL14(1–74). Each protein was assembled from three synthetic peptide fragments in “one-pot” using two sequential ligation reactions. We show that while glycosylation of CCL14(1–74) did not affect CCR1 binding affinity or potency of activation, sialylated variants of CCL14(1–74) exhibited reduced activity after treatment with plasmin compared to nonsialylated forms. These data indicate that glycosylation may influence the biological activity of CCL14 by regulating its conversion from the full-length to the truncated, activated form.

Citation

Wang, S., Foster, S. R., Sanchez, J., Corcilius, L., Larance, M., Canals, M., Stone, M. J., & Payne, R. J. (2021). Glycosylation Regulates N-Terminal Proteolysis and Activity of the Chemokine CCL14. ACS Chemical Biology, 16(6), 973-981. https://doi.org/10.1021/acschembio.1c00006

• Journal Article Type: Article
• Acceptance Date: May 3, 2021
• Online Publication Date: May 14, 2021
• Publication Date: Jun 18, 2021
• Deposit Date: Nov 19, 2021
• Journal: ACS Chemical Biology
• Print ISSN: 1554-8929
• Electronic ISSN: 1554-8937
• Publisher: American Chemical Society
• Peer Reviewed: Peer Reviewed
• Volume: 16
• Issue: 6
• Pages: 973-981
• DOI: https://doi.org/10.1021/acschembio.1c00006
• Keywords: Molecular Medicine; Biochemistry; General Medicine
• Public URL: https://nottingham-repository.worktribe.com/output/5536250
• Publisher URL: https://pubs.acs.org/doi/10.1021/acschembio.1c00006