Ligand specificity of purified complement receptor type three (CD11b/CD18, alpha m beta 2, Mac-1). Indirect effects of an Arg-Gly-Asp (RGD) sequence.
We have purified CR3 to homogeneity by affinity chromatography on C3bi-Sepharose and elution with EDTA. C3bi-coated erythrocytes bound to this purified CR3, and binding was dependent on the concentration of both C3bi and CR3, as well as on temperature and the presence of divalent cations. Moreover, binding could be blocked by mAb against CR3 or C3bi and could be enhanced by the addition of integrin modulating factor-1. We used the purified CR3 to test whether several putative ligands of CR3 directly bound the receptor. The interaction of purified CR3 with fibrinogen, filamentous hemagglutinin of Bordetella pertussis, lipophosphoglycan and glycoprotein 63 of Leishmania mexicana, and lipopolysaccharide from Escherichia coli was confirmed. However the interaction of CR3 with zymosan or its major component, beta-glucan, was not observed in these assays. Previous studies showed that binding of C3bi to PMN could be blocked with Arg-Gly-Asp (RGD) containing peptides and were interpreted to indicate that the RGD sequence in C3bi interacts directly with CR3. We found, however, that RGD containing peptides were unable to block the interaction of C3bi with purified CR3, yet retained the ability to block binding of C3bi to cells. We conclude that RGD-peptides do not directly bind CR3, but instead indirectly effect CR3 function. Inasmuch as the effect of RGD-peptides could be mimicked with antibodies against leukocyte response integrin, we suggest that RGD-peptides may bind to leukocyte response integrin on polymorphonuclear leukocytes and influence CR3 activity via this receptor.
Laboratory of Cellular Physiology and Immunology, Rockefeller University, New York, NY 10021.
J A
JA
Van Strijp
D G
DG
Russell
E
E
Tuomanen
E J
EJ
Brown
S D
SD
Wright
D000595:Amino Acid Sequence; D000818:Animals; D001667:Binding, Competitive; D002413:Cations, Divalent; D003179:Complement C3b / Q000378:metabolism; D005227:Fatty Acids / Q000378:metabolism; D005340:Fibrinogen / Q000378:metabolism; D006388:Hemagglutinins / Q000378:metabolism; D006801:Humans; D066298:In Vitro Techniques; D016023:Integrins / Q000378:metabolism; D007894:Leishmania mexicana / Q000378:metabolism; D008024:Ligands; D016177:Macrophage-1 Antigen / Q000302:isolation & purification / Q000378:metabolism*; D008969:Molecular Sequence Data; D009504:Neutrophils / Q000378:metabolism; D009842:Oligopeptides; D010455:Peptides / Q000378:metabolism
D002413:Cations, Divalent; D005227:Fatty Acids; D006388:Hemagglutinins; D016023:Integrins; D008024:Ligands; D016177:Macrophage-1 Antigen; D009842:Oligopeptides; D010455:Peptides; C072521:integrin modulating factor 1; C047981:arginyl-glycyl-aspartic acid; D003179:Complement C3b; D005340:Fibrinogen