The glycosylation properties of D2 dopamine receptors from striatal and limbic areas of bovine brain.
D2 dopamine receptors from bovine brain (caudate nucleus and olfactory tubercle) have been solubilized using sodium cholate/NaCl and their glycoprotein properties studied in terms of their interaction with wheat-germ agglutinin-agarose (WGA-agarose). Under optimal conditions about 65% of the applied D2 dopamine receptors bound to WGA-agarose and could be eluted with N-acetylglucosamine. The ability of receptors to adsorb to the affinity column was shown to be dependent on the cholate and salt concentrations used. Digestion of the membrane bound D2 dopamine receptors with neuraminidase prior to solubilisation reduced the ability of the receptors to bind to WGA-agarose (50% of applied receptors bound) whereas digestion with N-acetylglucosaminidase did not significantly affect binding to WGA-agarose. Digestion with the two enzymes together resulted in a larger decrease in binding to WGA-agarose than was seen with the two enzymes alone (40% of applied receptors bound). Stepwise elution of bound receptors from the WGA-agarose columns using 2.5 mM- and 100-mM-N-acetylglucosamine showed that about 40% of the bound receptors interacted with WGA-agarose in a low-affinity manner, the remainder showing a high-affinity interaction. Neuraminidase treatment reduced the low-affinity population suggesting that the interaction of oligosaccharides bearing sialic acid with WGA-agarose is of lower affinity and that higher-affinity binding is via N-acetylglucosamine. These data are discussed in terms of the heterogeneity of carbohydrate moieties on the D2 dopamine receptors within a brain region. In all the tests applied here, however, receptors from caudate nucleus and olfactory tubercle behaved identically so their glycosylation patterns must be very similar.
Leonard
M N
MN
Biological Laboratory, The University, Canterbury, Kent, U.K.
Williamson
R A
RA
Strange
P G
PG
D000118:Acetylglucosaminidase / Q000494:pharmacology; D000818:Animals; D001665:Binding Sites; D001921:Brain / Q000378:metabolism*; D002417:Cattle; D019826:Cholic Acid; D002793:Cholic Acids; D002846:Chromatography, Affinity; D004591:Electrophoresis, Polyacrylamide Gel; D006031:Glycosylation; D009439:Neuraminidase / Q000494:pharmacology; D011954:Receptors, Dopamine / Q000187:drug effects / Q000378:metabolism*; D017448:Receptors, Dopamine D2; D012965:Sodium Chloride; D013134:Spiperone / Q000494:pharmacology; D014018:Tissue Distribution
D002793:Cholic Acids; D011954:Receptors, Dopamine; D017448:Receptors, Dopamine D2; D012965:Sodium Chloride; D013134:Spiperone; D009439:Neuraminidase; D000118:Acetylglucosaminidase; D019826:Cholic Acid