Farooqui S M, Prasad C, Ali M
Laboratory of Neurosciences, Pennington Biomedical Research Center, Baton Rouge, LA 70808.
Biochem Biophys Res Commun. 1992 Apr 30;184(2):661-7. doi: 10.1016/0006-291x(92)90640-7.
A monoclonal antibody (Mab) that recognizes the rat dopamine D2 receptor (DAR) has been generated using DAR specific peptide. The Mab, IgM isotype recognizes five proteins (Mr 220, 145, 95, 66 and 47 kDa) in striatal membrane on Western blot. Preincubation of Mab with free peptide blocked the labeling of all five bands. A polyclonal antibody against peptide from a different region of the DAR, reacted with three out of five proteins (220, 66, and 47 kDa) in these membranes. The DAR antagonist NAPS-biotinyl binds to a 220 kDa protein in striatal membrane on ligand blotts; the labeling can be blocked by the addition of 2 microM sulpride. The 220 kDa Mab reactive protein was less in cerebellum and was absent in the liver. Neither the Mab nor polyclonal antibody inhibited binding of a DAR antagonist, [3H]YM09151-2, to the striatal membranes. These antibodies will enable us to study the structure/function and regulation of the synthesis of DAR protein.
利用多巴胺D2受体（DAR）特异性肽段制备了一种识别大鼠多巴胺D2受体的单克隆抗体（Mab）。该Mab为IgM同型，在蛋白质印迹法中可识别纹状体膜上的五种蛋白质（分子量分别为220、145、95、66和47 kDa）。用游离肽预先孵育Mab可阻断所有五条带的标记。一种针对DAR不同区域肽段的多克隆抗体，与这些膜中的五种蛋白质中的三种（220、66和47 kDa）发生反应。DAR拮抗剂NAPS-生物素在配体印迹中与纹状体膜上的一种220 kDa蛋白质结合；加入2 μM舒必利可阻断标记。220 kDa的Mab反应性蛋白在小脑中较少，在肝脏中不存在。Mab和多克隆抗体均未抑制DAR拮抗剂[3H]YM09151-2与纹状体膜的结合。这些抗体将使我们能够研究DAR蛋白的结构/功能以及合成调控。
