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

被引：15

作者：

Andersen, B ^{[1
]}

Stevens, RC

机构：

[1] Univ Calif Berkeley, Lawrence Berkeley Lab, Donner Lab, Div Life Sci, Berkeley, CA 94720 USA

[2] Royal Vet & Agr Univ, Dept Plant Biol, DK-1871 Frederiksberg C, Denmark

[3] Univ Calif Berkeley, Dept Chem, Berkeley, CA 94720 USA

来源：

PROTEIN EXPRESSION AND PURIFICATION | 1998年 / 13卷 / 01期

关键词：

D O I：

10.1006/prep.1998.0862

中图分类号：

Q5 [生物化学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：111 / 119

页数：9

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