亲和层析法分离大鼠脂肪细胞膜上腺苷A_1受体

Isolation of Adenosine A_1 Receptor from Rat Adipocyte Membrane

本文采用腺苷亲和层析法从大鼠脂肪细胞膜上分离出了一种亚基分子量为38kD的腺苷结合蛋白质。此蛋白在SDS-聚丙烯酰胺凝胶电泳上显示单一带,糖蛋白染色阳性;能与[8-~3H]腺苷特异结合(Kd=0.269nmol/L,Bmax=6.05pmol/mg.Pr);结合抑制实验表明它与腺苷A_1受体激动剂R-PIA、A_2受体激动剂NECA和腺苷的亲和力大小顺序为:R-PIA>腺苷>NECA。这表明所分离出的38kD蛋白是大鼠脂肪细胞膜上的腺苷A_1受体。

A protein, which exhibited a single band stained by Coomassie Blue and Schiff's Stain with an apparent Mr 38kD on SDS-PAGE, was purified from rat adipocyte membrane by affinity chromatography on a gel in which N^6-(6-aminohexyl) adenosine is covalently coupled to Affi-Gel 10. It could bind specifiealy with ~3H-adenosine, which was blocked by adenosine receptor agoaists R-PIA and NECA but not by adenosine transporter inhibitors. It is suggested that the glucoprotein purified here is the adenosine A_1receptor in rat adipoeyte membrane.