Insights into the Molecular Basis of Leukocyte Tethering and Rolling Revealed by Structures of P- and E-Selectin Bound to SLeX and PSGL-1

P-, E- and L-selectin constitute a family of cell adhesion receptors that mediate the initial tethering and rolling of leukocytes on inflamed endothelium as a prelude to their firm attachment and extravasation into tissues. The selectins bind weakly to sialyl Lewisx (SLe(X))-like glycans, but with high-affinity to specific glycoprotein counterreceptors, including PSGL-1. Here, we report crystal structures of human P- and E-selectin constructs containing the lectin and EGF (LE) domains co-complexed with SLe(X). We also present the crystal structure of P-selectin LE co-complexed with the N-terminal domain of human PSGL-1 modified by both tyrosine sulfation and SLe(X). These structures reveal differences in how E- and P-selectin bind SLe(X) and the molecular basis of the high-affinity interaction between P-selectin and PSGL-1.

[1]  G. Bricogne,et al.  [27] Maximum-likelihood heavy-atom parameter refinement for multiple isomorphous replacement and multiwavelength anomalous diffraction methods. , 1997, Methods in enzymology.

[2]  Kuo-Sen Huang,et al.  Insight into E-selectin/ligand interaction from the crystal structure and mutagenesis of the lec/EGF domains , 1994, Nature.

[3]  W. Weis,et al.  Structure of the calcium-dependent lectin domain from a rat mannose-binding protein determined by MAD phasing. , 1991, Science.

[4]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .

[5]  G S Kansas,et al.  Selectins and their ligands: current concepts and controversies. , 1996, Blood.

[6]  R. Cummings,et al.  Noncovalent Association of P-selectin Glycoprotein Ligand-1 and Minimal Determinants for Binding to P-selectin* , 2000, The Journal of Biological Chemistry.

[7]  T. Springer,et al.  Rolling and transient tethering of leukocytes on antibodies reveal specializations of selectins. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[8]  J. Bajorath,et al.  CD62/P-selectin binding sites for myeloid cells and sulfatides are overlapping. , 1994, Biochemistry.

[9]  B. Furie,et al.  Targeted Gene Disruption Demonstrates That P-Selectin Glycoprotein Ligand 1 (Psgl-1) Is Required for P-Selectin–Mediated but Not E-Selectin–Mediated Neutrophil Rolling and Migration , 1999, The Journal of experimental medicine.

[10]  Mary L. Westbrook,et al.  Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. , 1991 .

[11]  B. Brandley,et al.  Evidence for two classes of carbohydrate binding sites on selectins. , 1992, Glycobiology.

[12]  Brian Seed,et al.  PSGL-1 recognition of P-selectin is controlled by a tyrosine sulfation consensus at the PSGL-1 amino terminus , 1995, Cell.

[13]  R. Cummings,et al.  Role of PSGL-1 binding to selectins in leukocyte recruitment. , 1997, The Journal of clinical investigation.

[14]  S. Hemmerich,et al.  Sulfotransferases of Two Specificities Function in the Reconstitution of High Endothelial Cell Ligands for L-selectin , 1999, The Journal of cell biology.

[15]  K. Sharp,et al.  Protein folding and association: Insights from the interfacial and thermodynamic properties of hydrocarbons , 1991, Proteins.

[16]  R. Cummings,et al.  Perspectives series: cell adhesion in vascular biology. Role of PSGL-1 binding to selectins in leukocyte recruitment. , 1997, The Journal of clinical investigation.

[17]  G Bricogne,et al.  Direct phase determination by entropy maximization and likelihood ranking: status report and perspectives. , 1993, Acta crystallographica. Section D, Biological crystallography.

[18]  B. Furie,et al.  The Biology of P-Selectin Glycoprotein Ligand-1: Its Role as a Selectin Counterreceptor in Leukocyte-Endothelial and Leukocyte-Platelet Interaction , 1999, Thrombosis and Haemostasis.

[19]  A. Varki,et al.  Perspectives Series: Cell Adhesion in Vascular Biology Selectin Ligands: Will the Real Ones Please Stand Up? , 2022 .

[20]  K. Comess,et al.  A sulfated peptide segment at the amino terminus of PSGL-1 is critical for P-selectin binding , 1995, Cell.

[21]  Richard D. Cummings,et al.  Affinity and Kinetic Analysis of P-selectin Binding to P-selectin Glycoprotein Ligand-1* , 1998, The Journal of Biological Chemistry.

[22]  B. Furie,et al.  Expression cloning of a functional glycoprotein ligand for P-selectin , 1993, Cell.

[23]  M. U. Nollert,et al.  Tyrosine replacement in P-selectin glycoprotein ligand-1 affects distinct kinetic and mechanical properties of bonds with P- and L-selectin. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[24]  R. Cummings,et al.  The P-selectin glycoprotein ligand from human neutrophils displays sialylated, fucosylated, O-linked poly-N-acetyllactosamine. , 1994, The Journal of biological chemistry.

[25]  K. Comess,et al.  Isolated P-selectin Glycoprotein Ligand-1 Dynamic Adhesion to P- and E-selectin , 1997, Journal of Cell Biology.

[26]  A. Rehemtulla,et al.  Cloning and functional expression of a cDNA encoding the catalytic subunit of bovine enterokinase. , 1993, The Journal of biological chemistry.

[27]  T. Springer.  Traffic signals for lymphocyte recirculation and leukocyte emigration: The multistep paradigm , 1994, Cell.

[28]  John S. Philo,et al.  Conformation of sLex Tetrasaccharide, Free in Solution and Bound to E-, P-, and L-Selectin†,‡ , 1997 .

[29]  Richard D. Cummings,et al.  Tyrosine Sulfation of P-selectin Glycoprotein Ligand-1 Is Required for High Affinity Binding to P-selectin (*) , 1995, The Journal of Biological Chemistry.

[30]  Timothy A. Springer,et al.  The Kinetics of L-selectin Tethers and the Mechanics of Selectin-mediated Rolling , 1997, The Journal of cell biology.

[31]  Timothy A. Springer,et al.  Modifying the mechanical property and shear threshold of L-selectin adhesion independently of equilibrium properties , 1998, Nature.

[32]  D. Hammer,et al.  Lifetime of the P-selectin-carbohydrate bond and its response to tensile force in hydrodynamic flow , 1995, Nature.

[33]  E A Merritt,et al.  Raster3D Version 2.0. A program for photorealistic molecular graphics. , 1994, Acta crystallographica. Section D, Biological crystallography.

[34]  K. Croce,et al.  Interaction between soluble P-selectin and soluble P-selectin glycoprotein ligand 1: equilibrium binding analysis. , 1998, Biochemistry.

[35]  K. Drickamer,et al.  Two distinct classes of carbohydrate-recognition domains in animal lectins. , 1988, The Journal of biological chemistry.

[36]  D. Vestweber,et al.  Mechanisms that regulate the function of the selectins and their ligands. , 1999, Physiological reviews.

[37]  Wayne A. Hendrickson,et al.  Structure of a C-type mannose-binding protein complexed with an oligosaccharide , 1992, Nature.

[38]  A. Tulinsky,et al.  Structure of the hirugen and hirulog 1 complexes of alpha-thrombin. , 1991, Journal of molecular biology.

[39]  R. Cummings,et al.  Visualization of P-selectin Glycoprotein Ligand-1 as a Highly Extended Molecule and Mapping of Protein Epitopes for Monoclonal Antibodies (*) , 1996, The Journal of Biological Chemistry.

[40]  C. Bertozzi.  Cracking the carbohydrate code for selectin recognition. , 1995, Chemistry & biology.

[41]  S. Watson,et al.  The three members of the selectin receptor family recognize a common carbohydrate epitope, the sialyl Lewis(x) oligosaccharide , 1992, The Journal of cell biology.

[42]  A. Varki,et al.  Selectin ligands. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[43]  B. Honig,et al.  Classical electrostatics in biology and chemistry. , 1995, Science.

[44]  R. Cummings,et al.  A Novel Glycosulfopeptide Binds to P-selectin and Inhibits Leukocyte Adhesion to P-selectin* , 1999, The Journal of Biological Chemistry.

[45]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[46]  H. Erickson,et al.  Soluble monomeric P-selectin containing only the lectin and epidermal growth factor domains binds to P-selectin glycoprotein ligand-1 on leukocytes. , 1997, Blood.

[47]  R. Cummings,et al.  Structures of the O-Glycans on P-selectin Glycoprotein Ligand-1 from HL-60 Cells* , 1996, The Journal of Biological Chemistry.