The interaction of small oligomers of complement 3B (C3B) with phagocytes. High affinity binding and phorbol ester-induced internalization by polymorphonuclear leukocytes.
The binding of soluble, multimeric ligands of the major cleavage fragment of complement component 3 (C3b) to polymorphonuclear leukocytes (PMN) has been examined. The oligomers bound entirely via complement C3 receptor type 1 (CR1). There was a single affinity of binding (0.65 nM) at 37 degrees C, while this high affinity binding accounted for only a minority of ligand bound at 0 degree C, with the rest showing a 50-100-fold lower affinity. Azide, fluoride, cytochalasin B, and colchicine had no effect on oligomer binding to PMN. Binding of oligomers had no effect on CR1 expression by PMN. C3b oligomers were not spontaneously internalized by PMN, but were internalized in response to phorbol dibutyrate (PDBu). Both CR1 initially present on the PMN plasma membrane and CR1 initially present in the internal pool of receptors were able to participate in PDBu-induced ligand internalization. C3b oligomers attached to the detergent-insoluble cell cytoskeleton during incubation at 37 degrees C, but cytochalasin B did not inhibit PDBu-induced ligand internalization. Internalized ligand was no longer associated with the detergent-insoluble cytoskeleton. These data demonstrate that 1) some CR1 diffusion is required for optimal oligomer binding; 2) high affinity ligand is not a signal for plasma membrane expression of the internal pool of CR1; 3) CR1 cross-linking is not a sufficient signal for endocytosis; and 4) functional CR1 association with the cytoskeleton which occurs at the plasma membrane is not required for ligand internalization.
Brown
E J
EJ
Department of Medicine, Washington University School of Medicine, St. Louis, Missouri 63110.
D002413:Cations, Divalent / Q000494:pharmacology; D003179:Complement C3b / Q000378:metabolism*; D003572:Cytochalasins / Q000494:pharmacology; D003599:Cytoskeleton / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D006801:Humans; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D007700:Kinetics; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D009504:Neutrophils / Q000378:metabolism*; D015240:Phorbol 12,13-Dibutyrate / Q000494:pharmacology*; D011951:Receptors, Complement / Q000378:metabolism*; D013696:Temperature
D002413:Cations, Divalent; D003572:Cytochalasins; D011951:Receptors, Complement; D015240:Phorbol 12,13-Dibutyrate; D009240:N-Formylmethionine Leucyl-Phenylalanine; D003179:Complement C3b