Purification of an N-acetyl-D-glucosamine specific lectin (P.B.A.) from epidermal cell membranes of Pieris brassicae L.

We report the isolation and the purification of an N-acetyl-D-glucosamine specific lectin capable of agglutinating either fixed trypsinized rabbit erythrocytes or chitin particles. An agglutinin assay based on the affinity of this lectin for the chitin was devised with fluorescent particles of scorpion cuticle to measure lectin activity during purification steps. Lectin was isolated from epidermal cell membranes; its molecular weight was determined by gel filtration and polyacrylamide electrophoresis in sodium dodecyl sulfate. Mr was estimated to be 43,000. Lectin could be constituted by two subunits. Mr of which was estimated to be 23,000. The specificity of this lectin against N-acetyl-D-glucosamine and its oligomers suggests a possible role in the dynamics of these saccharides during the cuticle cycle.