Two proteins modulating transendothelial migration of leukocytes recognize novel carboxylated glycans on endothelial cells.
We recently showed that a class of novel carboxylated N:-glycans was constitutively expressed on endothelial cells. Activated, but not resting, neutrophils expressed binding sites for the novel glycans. We also showed that a mAb against these novel glycans (mAbGB3.1) inhibited leukocyte extravasation in a murine model of peritoneal inflammation. To identify molecules that mediated these interactions, we isolated binding proteins from bovine lung by their differential affinity for carboxylated or neutralized glycans. Two leukocyte calcium-binding proteins that bound in a carboxylate-dependent manner were identified as S100A8 and annexin I. An intact N terminus of annexin I and heteromeric assembly of S100A8 with S100A9 (another member of the S100 family) appeared necessary for this interaction. A mAb to S100A9 blocked neutrophil binding to immobilized carboxylated glycans. Purified human S100A8/A9 complex and recombinant human annexin I showed carboxylate-dependent binding to immobilized bovine lung carboxylated glycans and recognized a subset of mannose-labeled endothelial glycoproteins immunoprecipitated by mAbGB3.1. Saturable binding of S100A8/A9 complex to endothelial cells was also blocked by mAbGB3.1. These results suggest that the carboxylated glycans play important roles in leukocyte trafficking by interacting with proteins known to modulate extravasation.
Srikrishna
G
G
The Burnham Institute, La Jolla, CA 92037, USA.
Panneerselvam
K
K
Westphal
V
V
Abraham
V
V
Varki
A
A
Freeze
H H
HH
D000595:Amino Acid Sequence; D000818:Animals; D017305:Annexin A1 / Q000737:chemistry / Q000276:immunology / Q000378:metabolism; D000911:Antibodies, Monoclonal / Q000378:metabolism / Q000494:pharmacology; D000943:Antigens, Differentiation / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D002135:Calcium-Binding Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D040501:Calgranulin A; D040502:Calgranulin B; D002264:Carboxylic Acids / Q000378:metabolism*; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism* / Q000502:physiology; D002417:Cattle; D002448:Cell Adhesion / Q000276:immunology; D002462:Cell Membrane / Q000276:immunology / Q000378:metabolism; D002465:Cell Movement / Q000276:immunology; D002846:Chromatography, Affinity / Q000379:methods; D004730:Endothelium, Vascular / Q000166:cytology* / Q000276:immunology / Q000378:metabolism*; D006020:Glycopeptides / Q000138:chemical synthesis / Q000378:metabolism; D006801:Humans; D007106:Immune Sera / Q000378:metabolism / Q000494:pharmacology; D007962:Leukocytes / Q000276:immunology / Q000378:metabolism*; D008168:Lung / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009504:Neutrophils / Q000276:immunology / Q000378:metabolism; D011134:Polysaccharides / Q000378:metabolism*; D011817:Rabbits; D009418:S100 Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism / Q000502:physiology; D017386:Sequence Homology, Amino Acid
D017305:Annexin A1; D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D002135:Calcium-Binding Proteins; D040501:Calgranulin A; D040502:Calgranulin B; D002264:Carboxylic Acids; D002352:Carrier Proteins; D006020:Glycopeptides; D007106:Immune Sera; D011134:Polysaccharides; D009418:S100 Proteins