Akira Takeya

Isolation and Characterization of Anti-H Antibody from Egg Yolk of Immunized Hens

An anti-H antibody was demonstrated to be produced in egg yolk as well as in serum of hens which were immunized with human type O red blood cells. The antibody in egg yolk was isolated with polyethylene glycol and ethanol and was purified by affinity chromatography using immunoadsorbant beads immobilized with H type 2 hapten (Fuca1----2Gal beta 1----4GlcNAc beta). Hemagglutination reaction of the antibody from egg yolk was inhibited by human saliva samples from secretor types irrespective of their ABO blood types, and by immunoadsorbant beads which contain Fuca1----2Gal beta structures.

ウシとヒト乳汁中の(β1-3)N-アセチルグルコサミニルトランスフェラーゼとラクト-N-トリオースII構造をもつオリゴ糖の関係

We measured UDP-GlcNAc:Gal (beta 1-4) Glc (or GlcNAc) (beta 1-3) N-acetylglucosaminyltransferase activities in bovine (Holstein and Jersey cow) and human colostrums, and found in human colostrums sufficient activity to study the enzyme properties while not in bovine colostrums. The properties (requirements, pH optimum, acceptor specificity and Km values for lactose and N-acetyllactosamine) of the enzyme from human colostrum were very similar to those from human serum and urine. The reaction product was hydrolyzed by beta-N-acetylhexosaminidase, indicating that the N-acetylglucosaminyl residue was beta-linked to lactose. Methylation and hydrolysis of the reaction product from lactose [3H] labeled at the terminal galactose yielded 2, 4, 6-tri-O-methyl [3H] galactose. Thus the structure of the product was demonstrated to be GlcNAc (beta 1-3) Gal (beta 1-4) Glc (lacto-N-triose II). On the other hand, bovine sera contained N-acetylglucosaminyltransferase catalyzing the transfer of N-acetylglucosamine from UDP-GlcNAc to lactose. The enzyme activities were approximately 1/6-1/4 of that contained in human serum. The presence of (beta 1-3) N-acetylglucosaminyltransferase in human colostrum and its absence in bovine colostrums, apparently corresponds with the presence and absence of oligosaccharides containing lacto-N-triose II structure in colostrum.

A novel lectin in rabbit serum binds H type 1, H type 2 and N-acetyl lactosamine structures

A novel lectin (RSL) which recognizes blood group H type 1 and type 2 (Fuc alpha 1-->2Gal beta 1-->3/4GlcNAc beta-R), and N-acetyllactosamine (Gal beta 1-->4GlcNAc beta-R) was purified from rabbit serum using affinity chromatography on Synsorb H type 2 beads, gel filtration and preparative polyacrylamide gel electrophoresis. The lectin agglutinated human O type red cells, and the hemagglutination reaction was inhibited by H type 1 and type 2 haptens, N-acetyllactosamine and human salivas from secretor individuals. The molecular weight of the lectin was estimated to be approximate 650,000 and 65,000 on Sephacryl S-400 gel filtration and SDS-polyacrylamide gel electrophoresis, respectively.