Studies on the antigenicity and carbohydrates of human low molecular weight kininogen

Native low mol. wt (LMr) kininogen from human plasma and a kinin-free kininogen from Cohn's plasma fraction IV (HC-antigen) were isolated and studied with regard to their immunological reactivity and carbohydrate heterogeneity. Antisera were prepared against the conformational and sequence-dependent determinants of the heavy chain which is the common denominator of plasma kininogens. The molecules were characterized by single radial immunodiffusion and SDS gel electrophoresis. The effect of exoglycosidase treatment was investigated by a radioimmunoassay. It was found that the antigenic combining sites were not influenced by the partial removal of carbohydrates. These results suggest that the determinant structures of LMr kininogen reside in the peptide backbone of the protein. Carbohydrate heterogeneity was shown by different binding to concanavalin A and wheat germ lectin. With concanavalin A 52% of the immunoreactive HC-antigen was reactive compared to 76% of the native LMr kininogen. The weak reactivity (11%) of the HC-antigen towards the wheat germ lectin indicates that the N-acetylglucosamine residues are apparently blocked or missing. Immunoreactive kininogen was in all cases measured by single radial immunodiffusion or an RIA. The molecular nature of the heterogeneity is discussed in relation to the known isoelectric variations of kininogen.