The human D1A dopamine receptor:: Heterologous expression in Saccharomyces cerevisiae and purification of the functional receptor

Functional human D-1A dopamine receptor has been expressed in Saccharomyces cerevisiae. The primary sequence of the receptor was modified to include two affinity tags at the C-terminus of the protein, a FLAG tag (DYKDDDDK), and a His(6) tag (HHHHHH), These modifications allowed for purification to near homogeneity using immobilized metal affinity chromatography and immunoaffinity chromatography. Radioligand binding demonstrated that the purified and reconstituted receptor binds the antagonist [H-3]SCH23390 with an affinity (K-D = 8.0 +/- 3.2 nM) comparable to that of the native receptor. (C) 1998 Academic Press.