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Glycobiology Advance Access originally published online on February 29, 2008
Glycobiology 2008 18(6):441-446; doi:10.1093/glycob/cwn020
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© 2008 The Author(s)
This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.0/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

PSGL-1 from the murine leukocytic cell line WEHI-3 is enriched for core 2-based O-glycans with sialyl Lewis x antigen

Ziad S Kawar2,5, Thomas K Johnson3,5, Suvi Natunen4,5, John B Lowe6 and Richard D Cummings1,3,7

5 Department of Biochemistry and Molecular Biology, University of Oklahoma Health Sciences Center, Oklahoma City, OK 73104
6 Department of Pathology, Case Western Reserve University School of Medicine, Cleveland, OH 44106
7 Department of Biochemistry, Emory University School of Medicine, O. Wayne Rollins Research Center, 1510 Clifton Road, Suite 4001, Atlanta, GA 30322, USA

1 To whom correspondence should be addressed: Tel: +1-404-727-5962 (main office); Fax: +1-404-727-2738; e-mail: rdcummi{at}emory.edu

Received on January 17, 2008; revised on February 27, 2008; accepted on February 27, 2008

Leukocyte trafficking involves specific recognition between P-selectin and L-selectin and PSGL-1 containing core 2-based O-glycans expressing sialyl Lewis x (SLex) 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 SLex 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 3H-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 SLex 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 SLex antigen.

Key words: Lewis x / murine / O-glycan / P-selectin / PSGL-1

2 Selexys Pharmaceuticals Corp., 800 Research Parkway Suite, 338 Oklahoma City, OK 73104, USA.

3 Department of Biochemistry, Emory University School of Medicine, O. Wayne Rollins Research Center, 1510 Clifton Road, Suite 4001, Atlanta, GA 30322, USA.

4 Cell Surface Analytics Laboratory, Research and Development, Finnish Red Cross Blood Service, Kivihaantie 7, 00310 Helsinki, Finland.


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