Studies on the structure and carbohydrate binding properties of lobster agglutinin 1 (LAg1), a sialic acid-binding lectin.

Lobster agglutinin 1 (LAg 1) was isolated from the hemolymph of the American lobster (Homarus americanus) by a sequential combination of ammonium sulfate precipitation, preparative starch block electrophoresis, gel filtration and affinity chromatography on Sepharose-Fetuin and Sepharose-Colominic acid columns. Two types of protomeric structures with molecular weights of 700 and 500 Kilodaltons respectively were isolated. These molecules are composed of noncovalently held subunits with a molecular weight of 70 Kilodaltons. Analysis of preparations by double immunodiffusion, polyacrylamide gel electrophoresis and isoelectrofocusing indicates that the LAg 1 obtained was a single molecular species. Hemagglutination inhibition experiments indicated that the best inhibitors were bovine mucin, glycophorin, fetuin and human IgM in that order. The desialylated forms of some of these proteins still bound lectin, although to a lesser degree than their intact sialylated counterparts. Affinity chromatography experiments indicated that LAg 1 binds to N-acetylneuraminic acid, N-acetylglucosamine and N-acetylgalactosamine. LAg 1 does not contain sialic acid nor neuraminidase activity: oligosaccharides associated with it appear to be either of the oligomannosyl or biantennary type. The sialic acid binding specificity of this lectin was used to separate immature mouse thymocytes (low sialic acid content) from mature thymocytes (high sialic acid content).