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© The Rockefeller University Press, 0021-9525/1997//707 $5.00
The Journal of Cell Biology, Volume 136, Number 3, , 1997 707-716

Article

L-Selectin from Human, but Not from Mouse Neutrophils Binds Directly to E-Selectin



Olaf Zöllner*, Martin C. Lenter*, James E. Blanks*, Eric Borges*, Martin Steegmaier*, Hans-Günther Zerwes{ddagger}, and Dietmar Vestweber*

* Institute of Cell Biology, ZMBE, University of Münster, D-48149 Münster, Germany; and {ddagger} Sandoz Pharma Ltd., CH-4002 Basel, Switzerland

L-Selectin on neutrophils as well as inducible E- and P-selectin on endothelium are involved in the recruitment of neutrophils into inflamed tissue. Based on cell attachment assays, L-selectin was suggested to function as a carbohydrate presenting ligand for E- and P-selectin. However, previous affinity isolation experiments with an E-selectin–Ig fusion protein had failed to detect L-selectin among the isolated E-selectin ligands from mouse neutrophils. We show here that L-selectin from human neutrophils, in contrast to mouse neutrophils, can be affinity-isolated as a major ligand from total cell extracts using E-selectin–Ig as affinity probe. Binding of human L-selectin to E-selectin was direct, since purified L-selectin could be reprecipitated with E-selectin–Ig. Recognition of L-selectin was abolished by sialidase-treatment, required Ca2+, and was resistant to treatment with endoglycosidase F. Binding of L-selectin to a P-selectin–Ig fusion protein was not observed. In agreement with the biochemical data, the anti–Lselectin mAb DREG56 inhibited rolling of human neutrophils on immobilized E-selectin–Ig but not on P-selectin–Ig. No such inhibitory effect was seen with the anti–mouse L-selectin mAb MEL14 on mouse neutrophils. Rolling of E-selectin transfectants on purified and immobilized human L-selectin was inhibited by mAb DREG56. We conclude that L-selectin on human neutrophils is a major glycoprotein ligand among very few glycoproteins that can be isolated by an E-selectin affinity matrix. The clear difference between human and mouse L-selectin suggests that E-selectin–binding carbohydrate moieties are attached to different protein scaffolds in different species.

Abbreviations used in this paper: ESL, E-selectin ligand; PMN, polymorphonuclear granulocytes; PSGL, P-selectin glycoprotein ligand.

Address all correspondence to Dietmar Vestweber, Institute of Cell Biology, ZMBE, Technologiehof, Mendelstr. 11, D-48149 Münster, Germany. Tel.: 49 251 83 86 17. Fax: 49 251 83 86 16.


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