L-Selectin from Human, but Not from Mouse Neutrophils Binds Directly to E-Selectin
Open Access
- 10 February 1997
- journal article
- Published by Rockefeller University Press in The Journal of cell biology
- Vol. 136 (3), 707-716
- https://doi.org/10.1083/jcb.136.3.707
Abstract
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.Keywords
This publication has 55 references indexed in Scilit:
- The E-selectin Ligand-1 Is Selectively Activated in Chinese Hamster Ovary Cells by the α(1,3)-Fucosyltransferases IV and VIIPublished by Elsevier BV ,1996
- Structures of the O-Glycans on P-selectin Glycoprotein Ligand-1 from HL-60 CellsJournal of Biological Chemistry, 1996
- Neutrophils use both shared and distinct mechanisms to adhere to selectins under static and flow conditions.JCI Insight, 1995
- The P-selectin Glycoprotein Ligand Functions as a Common Human Leukocyte Ligand for P- and E-selectinsPublished by Elsevier BV ,1995
- P-selectin glycoprotein ligand-1 mediates rolling of human neutrophils on P-selectin.The Journal of cell biology, 1995
- Lymphocyte homing and leukocyte rolling and migration are impaired in L-selectin-deficient miceImmunity, 1994
- Traffic signals for lymphocyte recirculation and leukocyte emigration: The multistep paradigmCell, 1994
- Five tumor necrosis factor-inducible cell adhesion mechanisms on the surface of mouse endothelioma cells mediate the binding of leukocytes.The Journal of cell biology, 1993
- Identification of a glycoprotein ligand for E-selectin on mouse myeloid cells.The Journal of cell biology, 1993
- A cell-surface molecule involved in organ-specific homing of lymphocytesNature, 1983