PSGL-1 from the murine leukocytic cell line WEHI-3 is enriched for core 2-based O-glycans with sialyl Lewis x antigen.
Leukocyte trafficking involves specific recognition between P-selectin and L-selectin and PSGL-1 containing core 2-based O-glycans expressing sialyl Lewis x (SLe(x)) antigen. However, the structural identity of the glycan component(s) displayed by murine neutrophil PSGL-1 that contributes to its P-selectin counter-receptor activity has been uncertain, since these cells express little if any SLe(x) antigen, and because there have been no direct studies to examine murine PSGL-1 glycosylation. To address this uncertainty, we studied PSGL-1 glycosylation in the murine cell line WEHI-3 using metabolic-radiolabeling with (3)H-monosaccharide precursors to detect low-abundance O-glycan structures. We report that PSGL-1 from WEHI-3 cells expresses a di-sialylated core 2 O-glycan containing the SLe(x) antigen. This fucosylated O-glycan is scarce on PSGL-1 and essentially undetectable in total leukocyte glycoproteins from WEHI-3 cells. These results demonstrate that WEHI-3 cells selectively fucosylate PSGL-1 to generate functionally important core 2-based O-glycans containing the SLe(x) antigen.
Kawar
Ziad S
ZS
Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104, USA.
Johnson
Thomas K
TK
Natunen
Suvi
S
Lowe
John B
JB
Cummings
Richard D
RD
D000818:Animals; D002460:Cell Line; D002465:Cell Movement; D005643:Fucose / Q000378:metabolism*; D006031:Glycosylation; D019041:L-Selectin / Q000378:metabolism; D016256:Lewis X Antigen; D008562:Membrane Glycoproteins / Q000096:biosynthesis*; D051379:Mice; D009504:Neutrophils / Q000166:cytology / Q000378:metabolism*; D019007:P-Selectin / Q000378:metabolism; D046188:Protein Modification, Translational / Q000502:physiology*
D016256:Lewis X Antigen; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D019041:L-Selectin; D005643:Fucose