Chapot M P, Cervantes P, Kaveri S, Durieu-Trautmann O, Delavier-Klutchko C, Emorine L, Couraud P O, Strosberg A D
J Recept Res. 1987;7(1-4):1-15. doi: 10.3109/10799898709054976.
We have studied the molecular properties of avian beta 1-adrenergic receptor and human beta 2-adrenergic receptor. The turkey erythrocytes beta 1-receptor has been solubilized in active form by digitonin and has been purified to homogeneity by affinity chromatography followed by electroelution from polyacrylamide gel. The photoactivable ligand, iodocyanopindololdiazirine, labels specifically a major 45 kDa and minor 55 kDa polypeptide in turkey erythrocytes, whereas in A431, it labels two polypeptides of molecular weights 65 kDa and 55 kDa. Both types of receptors are N- and possibly O-glycosylated but the turkey beta 1 receptor has only complex carbohydrates whereas the human beta 2 receptor has in addition oligo mannosidic polysaccharidic moiety. Polyclonal and monoclonal antibodies were raised against the beta 1- and beta 2-adrenergic receptors. Polyclonal antibodies were found to mimic beta-adrenergic agonists by stimulating adenylate cyclase upon binding to the receptors. The monoclonal antibodies precipitated both intact and affinity labeled receptors which they also revealed on immunoblots.
我们研究了禽β1 - 肾上腺素能受体和人β2 - 肾上腺素能受体的分子特性。火鸡红细胞β1受体已通过洋地黄皂苷以活性形式溶解，并通过亲和色谱法纯化至同质，随后从聚丙烯酰胺凝胶中进行电洗脱。光活化配体碘氰吲哚二氮丙啶在火鸡红细胞中特异性标记一条主要的45 kDa多肽和一条次要的55 kDa多肽，而在A431细胞中，它标记分子量为65 kDa和55 kDa的两条多肽。这两种类型的受体均进行了N - 糖基化，可能还进行了O - 糖基化，但火鸡β1受体仅具有复合碳水化合物，而人β2受体还具有寡甘露糖苷多糖部分。制备了针对β1 - 和β2 - 肾上腺素能受体的多克隆抗体和单克隆抗体。发现多克隆抗体在与受体结合时通过刺激腺苷酸环化酶来模拟β - 肾上腺素能激动剂。单克隆抗体沉淀了完整的和亲和标记的受体，它们在免疫印迹上也显示出这些受体。
