Specific binding of glycosylated beta-galactosidase to bovine brain synaptic membrane.
The binding of a series of glycosylated beta-galactosidases to a fraction rich in synaptic membrane of bovine brain was examined. beta-galactosidase modified with p-aminophenyl beta-D-galactopyranoside (beta-D-Gal beta-gal) was found the most effective in binding to synaptic membrane, followed by that modified with beta-D-glucopyranoside, whereas the enzyme modified with p-aminophenyl derivatives of alpha-D-galactopyranoside, alpha-D-glucopyranoside, and alpha- and beta-L-fucopyranoside were found not to bind to the membrane. The binding was dependent on time, temperature, and pH; the maximal binding was obtained within 15 min at 4 degrees C and the optimal pH was approximately 4.0. The binding of beta-D-Gal beta-gal was inhibited by free p-aminophenyl beta-D-galactopyranoside and by the treatment of synaptic membrane with trypsin or phospholipase A2 or C. The equilibrium dissociation constant and the maximal concentration of binding sites were determined by Scatchard analysis to be 470 +/- 35 nM and 27.5 +/- 3.1 pmol/mg protein (n = 1). The results suggest that a specific binding site for the specified carbohydrates exists in synaptic membrane and is involved in the internalization of glycoconjugates into nerve terminals.
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D000818:Animals; D001230:Aspergillus; D001921:Brain / Q000166:cytology* / Q000201:enzymology; D050260:Carbohydrate Metabolism; D002417:Cattle; D005696:Galactosidases / Q000378:metabolism*; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D010741:Phospholipases A / Q000378:metabolism; D054467:Phospholipases A2; D013329:Structure-Activity Relationship; D013570:Synaptic Membranes / Q000201:enzymology*; D013696:Temperature; D014357:Trypsin / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism; D001616:beta-Galactosidase / Q000378:metabolism*
D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases; D005696:Galactosidases; D001616:beta-Galactosidase; D014357:Trypsin