Rostami A, Rabbani M, Mir-Mohammad-Sadeghi M
Department of Physiology and Pharmacology, School of Medicine, Zanjan University of Medical Sciences, Zanjan, Iran.
J Biomol Tech. 2010 Jul;21(2):92-6.
Glycosylation of the mu-opioid receptor may play an important role on its function. Using nested PCR, N53Q mutation was prepared in the N-terminal region of the rat mu-opioid receptor cDNA and cloned into the pcDNA3 vector. The plasmids containing the wild-type and mutated receptor cDNA were transfected into the COS-7 cells. Intracellular cAMP was measured in the morphine-treated and untreated transfected cells using an ELISA kit. Plasmid expression was evaluated using X-gal staining. Intracellular concentration of cAMP in the N53Q-mutated cells was not significantly different from the wild-type. The expression of the transfected plasmids was confirmed. Therefore, based on these results, it seems that glycosylation at the N53 site of the rat mu-opioid receptor does not influence the function of this receptor significantly.
μ-阿片受体的糖基化可能对其功能起重要作用。利用巢式PCR在大鼠μ-阿片受体cDNA的N端区域制备N53Q突变,并克隆到pcDNA3载体中。将含有野生型和突变型受体cDNA的质粒转染到COS-7细胞中。使用ELISA试剂盒在经吗啡处理和未处理的转染细胞中测量细胞内cAMP。使用X-gal染色评估质粒表达。N53Q突变细胞中cAMP的细胞内浓度与野生型无显著差异。转染质粒的表达得到证实。因此,基于这些结果,似乎大鼠μ-阿片受体N53位点的糖基化不会显著影响该受体的功能。