Studies on carbohydrate structure and immunological properties of human urinary kallikreins

Some studies on carbohydrates structure and immunological properties of human urinary kallikreins purified from the human urine of healthy men (HUK) were investigated. A general technique for fractionating asparagine-linked oligosaccharides of HUK was applied. This involves serial chromatographies on concanavalin A-Sepharose 4B and other lectins bound Agarose. Asparagine-linked main oligosaccharides of both active and inactive types of HUK were mixture of tri- or tetra-antennary complex oligosaccharide having GlcNAC beta 1-4Man beta residue (bisecting GlcNAc) (type I-2), bi-antennary complex oligosaccharide having core Fuc alpha 1-6GlcNAc residue (type III-3), tri- or tetra-antennary complex oligosaccharides having Gal beta 1-4GlcNAc beta 1-4 (Gal beta 1-4GlcNAc beta 1-2)Man alpha-residue (type I-1-A-a) etc. Besides these structural studies, the method to probe the heterogeneous profiles of HUK due to the different structures of carbohydrate chains bound, was devised without the complete purification of urinary kallikrein even with individual subject of not only active but also inactive forms of the kallikrein.