Changes in quaternary structure of IgG upon reduction of the interheavy-chain disulfide bond.

Abstract

Reduction of the single disulfide bond between H chains in the hinge region of rabbit Ig[immunoglobulin]G antibody [Ab] cause destabilization of the CH2 [H chain constant region 2] region of the molecule. Reduced Ab molecules may undergo a large change in quaternary structure in the CH2 region upon aggregation with a small bivalent hapten. The conformational change was observed in hydrodynamic studies and by EM. The sizes of native and reduced Ab complexes were measured from electron micrographs. These measurements show that reduction of the hinge disulfide allows the CH2 domains of the Ab to separate under the strain induced by complex formation. The Fab arms, which are clearly seen in the electron micrographs of the native complexes, are extended by a portion of the Fc region to effectively become Facb arms in the reduced complexes. The length of the arms is effectively increased by 23 .ANG.. This results in a massive alteration in the quaternary structure of the CH2 region of the molecule, and this may be the basis of many of the effects of mild reduction on the various effector functions of the Ab molecule. These findings support the open structure of the CH2 region proposed on the basis of crystallographic analyses, and they demonstrate how the inter-H chain hinge disulfide restricts segmental flexibility in the Fc fragment of the IgG molecule.