Iwanij V, Amos T M, Billington C J
Department of Genetics and Cell Biology, University of Minnesota, St. Paul 55108, USA.
Mol Cell Endocrinol. 1994 May;101(1-2):257-61. doi: 10.1016/0303-7207(94)90242-9.
125I-glucagon was directly cross-linked to its receptor in isolated adipocyte plasma membranes using a UV irradiation procedure. This investigation resulted in identification of an adipocyte glucagon receptor complex of 62 kDa, present both in white and brown adipose tissues. The specificity of labeling was shown by interference of unlabeled hormone with incorporation of radioactive glucagon into 62 kDa species. Treatment of adipose plasma membranes with N-glycanase resulted in appearance of intermediate species, indicating that the glucagon receptor is modified with several N-linked oligosaccharide chains similarly to the hepatic glucagon receptor. Peptide mapping of the affinity labeled adipose membranes with Staphylococcus aureus V8 protease generated three distinct receptor fragments identical to that of the hepatic receptor. Overall, the biochemical characterization of the rat adipocyte glucagon receptor indicates that it closely resembles the hepatic glucagon receptor.
使用紫外线照射程序将125I-胰高血糖素直接交联至分离的脂肪细胞质膜中的受体。这项研究鉴定出一种62 kDa的脂肪细胞胰高血糖素受体复合物,其存在于白色和棕色脂肪组织中。未标记激素对放射性胰高血糖素掺入62 kDa物质的干扰表明了标记的特异性。用N-聚糖酶处理脂肪细胞质膜会产生中间物种,这表明胰高血糖素受体与肝胰高血糖素受体类似,被多个N-连接的寡糖链修饰。用金黄色葡萄球菌V8蛋白酶对亲和标记的脂肪膜进行肽图谱分析,产生了与肝受体相同的三个不同的受体片段。总体而言,大鼠脂肪细胞胰高血糖素受体的生化特征表明它与肝胰高血糖素受体非常相似。