Biophys J, August 2000, p. 694-706, Vol. 79, No. 2

Sialyl Lewis^X-Mediated, PSGL-1-Independent Rolling Adhesion on P-selectin

Stephen D. Rodgers,^* Raymond T. Camphausen, and Daniel A. Hammer^*

 ^*Department of Chemical Engineering and Institute for Medicine and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania 19107, and  Genetics Institute, Cambridge, Massachusetts 02140 USA

Selectin-mediated cell adhesion is an essential component of the inflammatory response. In an attempt to unambiguously identify molecular features of ligands that are necessary to support rolling adhesion on P-selectin, we have used a reconstituted ("cell-free") system in which ligand-coated beads are perfused over soluble P-selectin surfaces. We find that beads coated with the saccharides sialyl Lewis^X (sLe^X), sialyl Lewis^a (sLe^a), and sulfated Lewis^X (HSO₃Le^X) support rolling adhesion on P-selectin surfaces. Although it has been suggested that glycosylation and sulfation of P-selectin glycoprotein ligand-1 (PSGL-1) is required for high-affinity binding and rolling on P-selectin, our findings indicate that sulfation of N-terminal tyrosine residues is not required for binding or rolling. However, beads coated with a tyrosine-sulfated, sLe^X-modified, PSGL-1-Fc chimera support slower rolling on P-selectin than beads coated with sLe^X alone, suggesting that sulfation improves rolling adhesion by modulating binding to P-selectin. In addition, we find it is not necessary that P-selectin carbohydrate ligands be multivalent for robust rolling to occur. Our results demonstrate that beads coated with monovalent sLe^X, exhibiting a more sparse distribution of carbohydrate than a similar amount of the multivalent form, are sufficient to yield rolling adhesion. The relative abilities of various ligands to support rolling on P-selectin are quantitatively examined among themselves and in comparison to human neutrophils. Using stop-time distributions, rolling dynamics at video frame rate resolution, and the average and variance of the rolling velocity, we find that P-selectin ligands display the following quantitative trend, in order of decreasing ability to support rolling adhesion on P-selectin: PSGL-1-Fc > sLe^a ~ sLe^X > HSO₃Le^X.

Biophys J, August 2000, p. 694-706, Vol. 79, No. 2
© 2000 by the Biophysical Society   0006-3495/00/08/694/13  $2.00

This article has been cited by other articles:

				H. Oh and S. L. Diamond Ethanol Enhances Neutrophil Membrane Tether Growth and Slows Rolling on P-Selectin but Reduces Capture from Flow and Firm Arrest on IL-1-Treated Endothelium J. Immunol., August 15, 2008; 181(4): 2472 - 2482. [Abstract] [Full Text] [PDF]

				C. D. Paschall, W. H. Guilford, and M. B. Lawrence Enhancement of L-Selectin, but Not P-Selectin, Bond Formation Frequency by Convective Flow Biophys. J., February 1, 2008; 94(3): 1034 - 1045. [Abstract] [Full Text] [PDF]

				L. A. Smith, H. Aranda-Espinoza, J. B. Haun, and D. A. Hammer Interplay between Shear Stress and Adhesion on Neutrophil Locomotion Biophys. J., January 15, 2007; 92(2): 632 - 640. [Abstract] [Full Text] [PDF]

				K. E. Edmondson, W. S. Denney, and S. L. Diamond Neutrophil-Bead Collision Assay: Pharmacologically Induced Changes in Membrane Mechanics Regulate the PSGL-1/P-Selectin Adhesion Lifetime Biophys. J., November 1, 2005; 89(5): 3603 - 3614. [Abstract] [Full Text] [PDF]

				A. O. Eniola, E. F. Krasik, L. A. Smith, G. Song, and D. A. Hammer I-Domain of Lymphocyte Function-Associated Antigen-1 Mediates Rolling of Polystyrene Particles on ICAM-1 under Flow Biophys. J., November 1, 2005; 89(5): 3577 - 3588. [Abstract] [Full Text] [PDF]

				K. E. Caputo and D. A. Hammer Effect of Microvillus Deformability on Leukocyte Adhesion Explored Using Adhesive Dynamics Simulations Biophys. J., July 1, 2005; 89(1): 187 - 200. [Abstract] [Full Text] [PDF]

				V. C. Ridger, P. G. Hellewell, and K. E. Norman L- and P-Selectins Collaborate to Support Leukocyte Rolling in Vivo When High-Affinity P-Selectin-P-Selectin Glycoprotein Ligand-1 Interaction Is Inhibited Am. J. Pathol., March 1, 2005; 166(3): 945 - 952. [Abstract] [Full Text] [PDF]

				S. Jadhav, C. D. Eggleton, and K. Konstantopoulos A 3-D Computational Model Predicts that Cell Deformation Affects Selectin-Mediated Leukocyte Rolling Biophys. J., January 1, 2005; 88(1): 96 - 104. [Abstract] [Full Text] [PDF]

				J. H. Im, W. Fu, H. Wang, S. K. Bhatia, D. A. Hammer, M. A. Kowalska, and R. J. Muschel Coagulation Facilitates Tumor Cell Spreading in the Pulmonary Vasculature during Early Metastatic Colony Formation Cancer Res., December 1, 2004; 64(23): 8613 - 8619. [Abstract] [Full Text] [PDF]

				E. F. Krasik and D. A. Hammer A Semianalytic Model of Leukocyte Rolling Biophys. J., November 1, 2004; 87(5): 2919 - 2930. [Abstract] [Full Text] [PDF]

				V. Ramachandran, M. Williams, T. Yago, D. W. Schmidtke, and R. P. McEver Dynamic alterations of membrane tethers stabilize leukocyte rolling on P-selectin PNAS, September 14, 2004; 101(37): 13519 - 13524. [Abstract] [Full Text] [PDF]

				W. D. Hanley, D. Wirtz, and K. Konstantopoulos Distinct kinetic and mechanical properties govern selectin-leukocyte interactions J. Cell Sci., May 15, 2004; 117(12): 2503 - 2511. [Abstract] [Full Text] [PDF]

				L. J. Rinko, M. B. Lawrence, and W. H. Guilford The Molecular Mechanics of P- and L-Selectin Lectin Domains Binding to PSGL-1 Biophys. J., January 1, 2004; 86(1): 544 - 554. [Abstract] [Full Text] [PDF]

				A. O. Eniola, P. J. Willcox, and D. A. Hammer Interplay between Rolling and Firm Adhesion Elucidated with a Cell-Free System Engineered with Two Distinct Receptor-Ligand Pairs Biophys. J., October 1, 2003; 85(4): 2720 - 2731. [Abstract] [Full Text] [PDF]

				S. K. Bhatia, M. R. King, and D. A. Hammer The State Diagram for Cell Adhesion Mediated by Two Receptors Biophys. J., April 1, 2003; 84(4): 2671 - 2690. [Abstract] [Full Text] [PDF]

				T. Yago, A. Leppanen, H. Qiu, W. D. Marcus, M. U. Nollert, C. Zhu, R. D. Cummings, and R. P. McEver Distinct molecular and cellular contributions to stabilizing selectin-mediated rolling under flow J. Cell Biol., August 19, 2002; 158(4): 787 - 799. [Abstract] [Full Text] [PDF]

				E. E. Burch, V. R. S. Patil, R. T. Camphausen, M. F. Kiani, and D. J. Goetz The N-terminal peptide of PSGL-1 can mediate adhesion to trauma-activated endothelium via P-selectin in vivo Blood, June 28, 2002; 100(2): 531 - 538. [Abstract] [Full Text] [PDF]

				M. R. King and D. A. Hammer Multiparticle adhesive dynamics: Hydrodynamic recruitment of rolling leukocytes PNAS, December 18, 2001; 98(26): 14919 - 14924. [Abstract] [Full Text] [PDF]