5′‐Nucleotidase from Bovine Caudate Nucleus Synaptic Plasma Membranes: Specificity for Substrates and Cations; Study of the Carbohydrate Moiety by Glycosidases

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Abstract
5''-Nucleotidase was studied in preparations of synaptic plasma membranes from bovine caudate nucleus. The best substrates for the membrane-bound enzyme were purine nucleotides, particularly 5''AMP. Effects of metal cations and chelating agents suggest that 5''-nucleotidase is a metalloprotein. Optimal conditions for solubilization of the 5''-nucleotidase were found by using a low concentration of the zwitterionic detergent sulfobetaine 14. Another membrane-bound enzyme, acetylcholinesterase, was not solubilized under these conditions, but only in the presence of Triton X-100. The effects of lectins (concanavalin A, Lens culinaris agglutinin, wheat germ agglutinin, and Limulus polyphemus agglutinin) showed that both enzymes are glycoproteins. Sequential hydrolysis with specific glycosidases produced modifications of the effect of lectins on these enzymes. The results suggest the presence of a complex-type glycosylation, with a fucose residue on the internal N-acetyl-D-glucosamine of the pentasaccharide core.