C1Q, THE COLLAGEN-LIKE SUBCOMPONENT OF THE 1ST COMPONENT OF COMPLEMENT C1, IS A MEMBRANE-PROTEIN OF GUINEA-PIG MACROPHAGES

C1q, a subcomponent of C1 - the first component of complement, is synt hesized by macrophages (MPHI). Immunofluorescence and immunoperoxidase studies first indicated the presence of C1q on the surface of guinea pig (gp) and human peritoneal MPHI (Loos, M., Storz, R., Muller, W. an d Lemmel, E.M., Immunobiology 1981, 158: 213). In our study different methods for labeling of gp serum and gp MPHI C1q were employed. The pr esence of C1q protein on the surface of gp peritoneal MPHI is shown by cell surface labeling with the biotin derivative sulfosuccinimdyl-6-( biotinamido)-hexanoate and subsequent immunoprecipitation. The mechani sm by which C1q is attached to the cell membrane was also investigated . Intact cells were treated with acid stripping-buffers or phosphatidy linositol-specific phospholipase C and separated membranes were extrac ted with a buffer containing 1 M KCl and 3 M urea. Regardless of which method was used, C1q remained attached to the membrane. When surface- labeled cells were cultured, they were found to release the C1q from t heir surface membrane into the culture medium. Lysates of biosynthetic ally labeled cells were used to show that, like secreted or serum C1q, cellular MPHI C1q binds to immobilized homologous IgG. This implies t hat the globular regions of the cellular C1q are functionally active. The results reveal that (i) cellular MPHI C1q is firmly located in the membrane throughout the biosynthetic pathway, such that it is compara ble with an integral membrane protein, (ii) cellular MPHI C1q is not r eversibly bound to the cell surface via a receptor. We suggest that C1 q, as a membrane protein of MPHI, serves as an Fc binding factor that also is secreted into the environment.