Purification of the B lymphocyte receptor for the C3d fragment of complement and the Epstein-Barr virus by monoclonal antibody affinity chromatography, and assessment of its functional capacities.
The human C3d receptor (complement receptor type 2, CR2), that also serves as the B lymphocyte receptor for the Epstein-Barr virus, was purified from detergent lysates from the B lymphoblastoid cell lines, SB and Raji, by monoclonal antibody affinity chromatography using the anti-CR2 monoclonal antibody, HB-5. Relative to the concentration of cellular protein and receptor that was initially solubilized by detergent, the procedure provided a 37,000-fold purification with a 40-50% recovery of CR2. The purified receptor presented a single Coomassie blue-stained band when analyzed by SDS-PAGE, and it retained its function of binding to C3-Sepharose. The N-terminus of CR2 was blocked. The amino acid composition was significantly similar to that of the C3b/C4b receptor, factor H and C4 binding protein, suggesting that CR2 may be a member of this newly defined protein family. However, CR2 did not exhibit the regulatory functions of these proteins, namely, the decay dissociation of the classical or alternative pathway C3 convertases and serving as a cofactor for the cleavage of C3b.
D000596:Amino Acids / Q000032:analysis; D000911:Antibodies, Monoclonal; D001402:B-Lymphocytes / Q000032:analysis* / Q000276:immunology; D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods; D003176:Complement C3 / Q000378:metabolism*; D017244:Complement Factor I; D010450:Endopeptidases / Q000502:physiology; D004854:Herpesvirus 4, Human; D006801:Humans; D008970:Molecular Weight; D011863:Radioimmunoassay; D011951:Receptors, Complement / Q000302:isolation & purification* / Q000502:physiology; D017464:Receptors, Complement 3d; D011991:Receptors, Virus / Q000032:analysis* / Q000502:physiology
D000596:Amino Acids; D000911:Antibodies, Monoclonal; D003176:Complement C3; D011951:Receptors, Complement; D017464:Receptors, Complement 3d; D011991:Receptors, Virus; D010450:Endopeptidases; D017244:Complement Factor I