Purification of brain D2 dopamine receptor.
D2 dopamine receptors have been extracted from bovine brain using the detergent cholate and purified approximately 20,000-fold by affinity chromatography on haloperidol-sepharose and wheat germ agglutinin-agarose columns. The purified preparation contains D2 dopamine receptors as judged by the pharmacological specificity of [3H]spiperone binding to the purified material. The sp. act. of [3H]spiperone binding in the purified preparation is 2.5 nmol/mg protein. The purified preparation shows a major diffuse band at Mr 95,000 upon SDS-polyacrylamide gel electrophoresis and there is evidence for microheterogeneity either at the protein or glycosylation level. Photoaffinity labelling of D2 dopamine receptors also shows a species of Mr 95,000. The D2 dopamine receptor therefore is a glycoprotein of Mr 95,000.
Williamson
R A
RA
Biological Laboratory, The University, Canterbury, Kent, UK.
Worrall
S
S
Chazot
P L
PL
Strange
P G
PG
D000345:Affinity Labels; D000818:Animals; D002417:Cattle; D002421:Caudate Nucleus / Q000032:analysis*; D002846:Chromatography, Affinity; D008970:Molecular Weight; D011954:Receptors, Dopamine / Q000302:isolation & purification*; D012995:Solubility; D013134:Spiperone / Q000031:analogs & derivatives
D000345:Affinity Labels; D011954:Receptors, Dopamine; C050343:4-azido-N-methylspiperone; D013134:Spiperone