Interaction between complement proteins C5b-7 and erythrocyte membrane sialic acid.
The initial phase of membrane attack by complement is the interaction between C5b6, C7, and the cell membrane that leads to the insertion of C5b-7. Here we investigate the role of sialic acid residues in the assembly of C5b-7 intermediates on erythrocyte cell membranes. We find that C5b6 binds to glycophorin, whereas C5 or C6 does not bind, and desialylation of the glycophorin abolishes C5b6 binding. Complement lysis is inhibited by either masking glycophorin sialic acid with F(ab) fragments of an mAb, or by removal of the sialylated region of glycophorin by mild trypsinization. Gangliosides inhibit C5b-7 deposition when added to the aqueous phase. Asialogangliosides and synthetic gangliosides lacking the carboxylic acid residue have no inhibitory activity. We conclude that C5b6 binds to sialylated molecules on the erythrocyte surface. We propose a new model of membrane attack in which C5b6 initially binds to membranes via ionic forces. C7 then binds to C5b6, disrupting the ionic interaction and leading to the exposure of hydrophobic domains. Sialic acid is known to inhibit complement activation. Thus, these findings reveal a paradoxical role for sialic acid in complement attack; the presence of sialic acid inhibits the generation of C5b6, but once the membrane attack pathway is initiated, sialic acid enhances complement lysis.
Marshall
P
P
Dermatology Service, Veterans Affairs Medical Center, New York 10016, USA.
Hasegawa
A
A
Davidson
E A
EA
Nussenzweig
V
V
Whitlow
M
M
D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001212:Asialoglycoproteins / Q000302:isolation & purification / Q000378:metabolism; D002240:Carbohydrate Sequence; D002846:Chromatography, Affinity; D003182:Complement C5 / Q000378:metabolism*; D015938:Complement Membrane Attack Complex / Q000378:metabolism; D004910:Erythrocyte Membrane / Q000378:metabolism*; D005679:G(M3) Ganglioside / Q000378:metabolism*; D005732:Gangliosides; D006021:Glycophorins / Q000187:drug effects / Q000302:isolation & purification / Q000378:metabolism*; D006168:Guinea Pigs; D006461:Hemolysis / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D019158:N-Acetylneuraminic Acid / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D001212:Asialoglycoproteins; D003182:Complement C5; D015938:Complement Membrane Attack Complex; D005679:G(M3) Ganglioside; D005732:Gangliosides; D006021:Glycophorins; C026503:asialoglycophorin; C037453:complement C5b-7 complex; D014357:Trypsin; D019158:N-Acetylneuraminic Acid