Interaction of P-selectin (CD62) and its cellular ligand: analysis of critical residues.
P-Selectin (CD62, PADGEM, GMP140) is a membrane glycoprotein which is rapidly mobilized to the surface of activated platelets and endothelial cells where it mediates leukocyte-platelet and leukocyte-vascular endothelial cell adhesion, respectively. P-Selectin is a member of a family of adhesion molecules which includes the endothelial cell adhesion molecule E-selectin and the leukocyte adhesion molecule L-selectin. Selectins mediate cell-cell binding resulting from the interaction between the amino terminal lectin domains of the selectins and their respective carbohydrate ligands. Here we report on a three-dimensional model of the lectin domain of P-selectin which was derived on the basis of its structural homology to the rat mannose binding protein (MBP) whose crystal structure has recently been reported. On the basis of the model, a number of point mutants were prepared to identify the P-selectin binding site. The residues found to be important for binding are located in a shallow groove on the surface of the molecule composed of residues from the beta-2, -3, and -5 strands of the P-selectin lectin domain. A number of residues within this groove, which are conserved among all selectins, were found to be critical for P-selectin binding. They include Lys113, Tyr48, and Tyr94. The single substitutions Lys113Ala, Tyr48Ala, Tyr48Phe, Tyr94Ala, and Tyr94Phe abolished P-selectin binding to myeloid cells.
Hollenbaugh
D
D
Bristol-Myers Squibb Pharmaceutical Research Institute, Seattle, Washington 98121.
Bajorath
J
J
Stenkamp
R
R
Aruffo
A
A
D000595:Amino Acid Sequence; D000818:Animals; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism; D002352:Carrier Proteins / Q000737:chemistry; D002448:Cell Adhesion; D003460:Crystallization; D006801:Humans; D037102:Lectins / Q000378:metabolism; D007962:Leukocytes / Q000378:metabolism; D037241:Mannose-Binding Lectins; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016296:Mutagenesis; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D016133:Polymerase Chain Reaction; D017433:Protein Structure, Secondary; D051381:Rats; D017386:Sequence Homology, Amino Acid; D014407:Tumor Cells, Cultured
D002352:Carrier Proteins; D037102:Lectins; D037241:Mannose-Binding Lectins; D019007:P-Selectin; D010980:Platelet Membrane Glycoproteins; D002118:Calcium