Labile disulfide bonds and free thiol groups in human IgG. I. Assignment to IgG1 and IgG2 subclasses

The content of free SH groups (about 0.24) and labile S-S bonds (about 0.64) per mole human IgG can be differentially determined by the reaction with 5,5'-dithio(2,2'-dinitro)benzoate (DTNB) for 30 min and 24 h, respectively. Highly significant linear correlations were found between the number of labile S-S and the percentage of IgG1, and the number of free SH and the percentage of IgG2 of the total IgG fraction. It is concluded that the IgG1 molecule contains one S-S bond which is opened during the 24 h interaction with DTNB by a disulfide exchange reaction. This was also confirmed by the investigation of pure monomeric IgG1. The splitting of this bond does not alter molecular weight, antigenic properties or antigen binding activity, but reduces significantly the complement binding activity of IgG. On the other hand, one free SH group could be found in the IgG2 molecule. Changes have been observed in SH and S-S levels of total IgG in patients with various malignant diseases that are to be explained by corresponding changes of the percentages of IgG1 and IgG2.