title: Identification of the Raji cell membrane-derived C1q inhibitor as a receptor for human C1q. Purification and immunochemical characterization.
abstract: We have shown previously that an activity which is capable of precipitating purified C1q and inhibiting some of the C1q-dependent biologic reactions could be solubilized from the membranes of both normal human peripheral B lymphocytes and a B cell-derived lymphoblastoid cell line (Raji), both of which are known to possess receptors for human C1q. In this report we present evidence that this membrane-associated C1q inhibitor is a chondroitinase-insensitive macromolecule and is the receptor for human C1q. The receptor was solubilized from membranes of Raji cells with Nonidet P-40 and purified to homogeneity using C1q-Sepharose 4B affinity chromatography. Equilibrium density gradient centrifugation analysis revealed that the complex could be resolved into a protein-rich, low density fraction and a carbohydrate-rich, high density fraction. The large hydrodynamic size, coupled with the high buoyant density, suggests that a proteoglycan is a constituent of the complex and indicates that the receptor might be a macromolecular complex of a proteoglycan portion noncovalently linked to a 60-70 kD glycoprotein. The glycoprotein moiety, in turn, consists of two or more identical (70,000 mol wt) polypeptide chains held together by disulfide bonds and constitutes the C1q receptor (C1qR). Sucrose density ultracentrifugation analysis showed that the isolated receptor sediments with an apparent rate of 4.2 S. Immunochemical analyses demonstrated that a typical preparation of the C1qR complex consists of approximately 23% uronic acid and approximately 21% galactosamine with a galactosamine-to-glucosamine ratio of 3.2. Binding of C1q to the receptor was found to be optimal at low ionic strength and neutral or near-neutral pH (7-7.4). The isolated receptor was found to inhibit C1q hemolytic function, abrogate C1q-dependent rosette formation, and block the C1q-dependent, cell-mediated cytotoxicity, all of which are activities mediated by the receptor.
mesh_terms: D000596:Amino Acids / Q000032:analysis; D001667:Binding, Competitive; D002051:Burkitt Lymphoma / Q000276:immunology; D002352:Carrier Proteins; D002460:Cell Line; D002499:Centrifugation, Density Gradient; D002846:Chromatography, Affinity; D003166:Complement Activating Enzymes / Q000276:immunology* / Q000378:metabolism; D015922:Complement C1q; D003169:Complement Inactivator Proteins / Q000502:physiology*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D018960:Hyaluronan Receptors; D005779:Immunodiffusion; D008562:Membrane Glycoproteins; D008565:Membrane Proteins / Q000378:metabolism*; D024101:Mitochondrial Proteins; D011951:Receptors, Complement / Q000032:analysis / Q000302:isolation & purification* / Q000502:physiology
chemical_list: D000596:Amino Acids; C087299:C1QBP protein, human; D002352:Carrier Proteins; D003169:Complement Inactivator Proteins; D018960:Hyaluronan Receptors; D008562:Membrane Glycoproteins; D008565:Membrane Proteins; D024101:Mitochondrial Proteins; D011951:Receptors, Complement; C032402:complement 1q receptor; D015922:Complement C1q; D003166:Complement Activating Enzymes