O-glycans direct selectin ligands to lipid rafts on leukocytes

Bojing Shao, Tadayuki Yago, Hendra Setiadi, Ying Wang, Padmaja Mehta-D'Souza, Jianxin Fu, Paul R. Crocker, William Rodgers, Lijun Xia, Rodger P. McEver (Lead / Corresponding author)

Research output: Contribution to journalArticle

21 Citations (Scopus)

Abstract

Palmitoylated cysteines typically target transmembrane proteins to domains enriched in cholesterol and sphingolipids (lipid rafts). P-selectin glycoprotein ligand-1 (PSGL-1), CD43, and CD44 are O-glycosylated proteins on leukocytes that associate with lipid rafts. During inflammation, they transduce signals by engaging selectins as leukocytes roll in venules, and they move to the raft-enriched uropods of polarized cells upon chemokine stimulation. It is not known how these glycoproteins associate with lipid rafts or whether this association is required for signaling or for translocation to uropods. Here, we found that loss of core 1-derived O-glycans in murine C1galt1<sup>-/-</sup> neutrophils blocked raft targeting of PSGL-1, CD43, and CD44, but not of other glycosylated proteins, as measured by resistance to solubilization in nonionic detergent and by copatching with a raft-resident sphingolipid on intact cells. Neuraminidase removal of sialic acids from wild-type neutrophils also blocked raft targeting. C1galt1<sup>-/-</sup> neutrophils or neuraminidase-treated neutrophils failed to activate tyrosine kinases when plated on immobilized anti-PSGL-1 or anti-CD44 F(ab′)<inf>2</inf>. Furthermore, C1galt1<sup>-/-</sup> neutrophils incubated with anti-PSGL-1 F(ab′)<inf>2</inf> did not generate microparticles. In marked contrast, PSGL-1, CD43, and CD44 moved normally to the uropods of chemokine-stimulated C1galt1<sup>-/-</sup> neutrophils. These data define a role for core 1-derived O-glycans and terminal sialic acids in targeting glycoprotein ligands for selectins to lipid rafts of leukocytes. Preassociation of these glycoproteins with rafts is required for signaling but not for movement to uropods.

Original languageEnglish
Pages (from-to)8661-8666
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume112
Issue number28
DOIs
Publication statusPublished - 14 Jul 2015

Keywords

  • Cell adhesion
  • Cell signaling
  • Inflammation
  • Neutrophil
  • Uropod

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  • Cite this

    Shao, B., Yago, T., Setiadi, H., Wang, Y., Mehta-D'Souza, P., Fu, J., Crocker, P. R., Rodgers, W., Xia, L., & McEver, R. P. (2015). O-glycans direct selectin ligands to lipid rafts on leukocytes. Proceedings of the National Academy of Sciences of the United States of America, 112(28), 8661-8666. https://doi.org/10.1073/pnas.1507712112