Studies on the combining sites of concanavalin A.

The initial event in the biological activity of concanavalin A (Con A) involves binding of the protein to cell surface receptors. The nature and mechanism whereby such binding may occur is described in terms of cell surface carbohydrates and the demonstrated specificity of the protein. Although considerable latitude is tolerated at the C-2 position of the alpha-D-hexopyranose ring system, the carbohydrate binding site of Con A appears to be complemnetary to alpha-D-mannopyranosyl residues. Hapten inhibition studies indicate that each of the hydroxyl groups of this sugar is probably involved in the binding mechanism. Of the common sugars present on cell surfaces (D-glucose, D-mannose and N-acetyl-D-glucosamine), it is probably alpha-D-mannopyranosyl residues which react with Con A. Since the latter units are primary receptors for Con A. Evidence supporting this view includes hapten inhibition studies with model oligosaccarides and preciptin studies with macromolecules containing internal 2-o-substituted alpha-D-mannopyranosyl residues. The binding to Con A of a series of oligosaccharides containing alpha-(1leads to2)-linked D-mannosyl units appears to increase up to the tetraose and then decreases; several possible explanations are considered. Acetylated Con A, although retaining its specificity, is about 50% as active as the native protein. Some biological properties of the modified protein are described. Data suggesting that Con A behaves differently in the solution phase than in the crystalline state are presented in terms of UV difference displacement studies. It is suggested that the so-called carbohydrate binding site reportedly identified in Con A crystals may not be correct.