Cutting edge: evidence for a signaling partnership between urokinase receptors (CD87) and L-selectin (CD62L) in human polymorphonuclear neutrophils.
Leukocyte urokinase plasminogen activator receptors (uPARs) cluster at adhesion interfaces and at migratory fronts where they participate in adhesion, chemotaxis, and proteolysis. uPAR aggregation triggers activation signaling even though this glycolipid-anchored protein must associate with membrane-spanning proteins to access the cell interior. This study demonstrates a novel partnership between uPAR and L-selectin in human polymorphonuclear neutrophils. Fluorescence resonance energy transfer demonstrated a direct physical association between uPAR and L-selectin. To examine the role of L-selectin in uPAR-mediated signaling, uPAR was cross-linked and intracellular Ca(2+) concentrations were measured by spectrofluorometry. A mAb reactive against the carbohydrate binding domain (CBD) of L-selectin substantially inhibited uPAR-mediated Ca(2+) mobilization, whereas mAbs against the beta(2) integrin complement receptor 3 (CR3), another uPAR-binding adhesion protein, had no effect. Similarly, fucoidan, a sulfated polysaccharide that binds to L-selectin CBD, inhibited the Ca(2+) signal. We conclude that uPAR associates with the CBD region of L-selectin to form a functional signaling complex.
Sitrin
R G
RG
Pulmonary and Critical Care Medicine Division, Department of Internal Medicine, University of Michigan, Ann Arbor MI 48109, USA. rsitrin@umich.edu
Pan
P M
PM
Blackwood
R A
RA
Huang
J
J
Petty
H R
HR
D050260:Carbohydrate Metabolism; D002448:Cell Adhesion / Q000276:immunology; D004735:Energy Transfer / Q000276:immunology; D006031:Glycosylation; D006801:Humans; D019041:L-Selectin / Q000276:immunology / Q000378:metabolism / Q000502:physiology*; D008024:Ligands; D018375:Neutrophil Activation / Q000276:immunology; D009504:Neutrophils / Q000201:enzymology / Q000276:immunology* / Q000378:metabolism*; D011485:Protein Binding / Q000276:immunology; D017434:Protein Structure, Tertiary; D011956:Receptors, Cell Surface / Q000502:physiology*; D055293:Receptors, Urokinase Plasminogen Activator; D015398:Signal Transduction / Q000276:immunology*; D013050:Spectrometry, Fluorescence; D014568:Urokinase-Type Plasminogen Activator / Q000378:metabolism*
D008024:Ligands; C524898:PLAUR protein, human; D011956:Receptors, Cell Surface; D055293:Receptors, Urokinase Plasminogen Activator; D019041:L-Selectin; D014568:Urokinase-Type Plasminogen Activator