The hinge as a spacer contributes to covalent assembly and is required for function of IgG

Abstract

Chimeric IgG3 lacking a genetic hinge but with Cys residues in CH2 between position 231 and 232 (IgG3 deltaH+Cys) and position 279 (IgG3 deltaH+2Cys) produced by site-directed mutagenesis have been found to be deficient in their intermolecular assembly. Although some H2L2 molecules are formed, significant quantities of assembly intermediates, in particular HL, are found in the secretions. Both IgG3 deltaH+Cys and IgG3 deltaH+2Cys were greatly reduced in their ability to bind Fc gammaRI. They also failed to bind C1q and activate the complement cascade. Addition of the tailpiece from IgM (microtp) to IgG3 deltaH+Cys (yielding IgG3 deltaH+Cys microtp) failed to lead to efficient oligomerization. IgG3 deltaH+Cys microtp failed to bind Fc gammaRII. However, at high concentrations it was able to bind C1q and effect complement-dependent cytolysis. It consumed complement in the absence of added Ag, indicating that removing the flexible hinge was not sufficient to yield polymeric IgG that resembled IgM in its dependence on added Ag for complement activation.