GMP-140 binding to neutrophils is inhibited by sulfated glycans

• Published in: The Journal of biological chemistry 1991-03, Vol.266 (9), p.5371-5374
• Main Authors: Skinner, M P, Lucas, C M, Burns, G F, Chesterman, C N, Berndt, M C
• Format: Article
• Language: English
• Subjects: Analytical, structural and metabolic biochemistry; Antibodies, Monoclonal - immunology; Biological and medical sciences; Cell Adhesion Molecules - immunology; Cell Adhesion Molecules - metabolism; Chromatography, Gel; endothelium; Fundamental and applied biological sciences. Psychology; Glycoproteins; Humans; leukocytes (neutrophilic); Neutrophils - immunology; Neutrophils - metabolism; P-Selectin; Platelet Membrane Glycoproteins - antagonists & inhibitors; Platelet Membrane Glycoproteins - immunology; Platelet Membrane Glycoproteins - metabolism; Polysaccharides - pharmacology; Proteins; Rosette Formation; Sulfates
• ISSN: 0021-9258; 1083-351X
• DOI: 10.1016/S0021-9258(19)67603-9

GMP-140 is a 140-kDa granule membrane glycoprotein localized to the alpha-granules of platelets and the Weibel-Palade bodies of endothelial cells. Expression of GMP-140 on the activated cell surface has been shown to mediate the adhesion of thrombin-activated platelets to neutrophils and monocytes and the transient adhesion of neutrophils to endothelium. In contrast, fluid-phase GMP-140 strongly inhibits the CD18-dependent adhesion of tumor necrosis factor alpha-activated neutrophils to endothelium suggesting that GMP-140 can also serve an anti-adhesive function. In the present report, it is demonstrated that fluid-phase GMP-140 which exists predominantly as a tetramer binds to a single class of high affinity receptor on neutrophils and HL60 cells. Binding of 125I-labeled GMP-140 to neutrophils and HL60 cells and the rosetting of neutrophils and HL60 cells by thrombin-activated platelets were inhibited by EDTA, excess unlabeled fluid-phase GMP-140, Fab fragments of an affinity-purified rabbit anti-GMP-140 antibody, and by the murine anti-GMP-140 monoclonal antibody, AK 4. Both neutrophil and HL60 GMP-140 binding and platelet rosetting were strongly inhibited by heparin, fucoidin, and dextran sulfate 500,000, were partially inhibited by dextran sulfate 5,000 and lambda- and kappa-carrageenan, but were not inhibited by chondroitins 4- and 6-sulfate. Since this sulfated glycan specificity is identical to that previously reported by us for GMP-140, the present results suggest that the sulfated glycan binding site and the neutrophil receptor binding site on GMP-140 are either identical or proximal.