首页> 外文期刊>Journal of Neuroimmunology: Official Bulletin of the Research Committee on Neuroimmunology of the World Federation of Neurology >Characterization of membrane melatonin receptor in mouse peritoneal macrophages: inhibition of adenylyl cyclase by a pertussis toxin-sensitive G protein.
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Characterization of membrane melatonin receptor in mouse peritoneal macrophages: inhibition of adenylyl cyclase by a pertussis toxin-sensitive G protein.

机译:小鼠腹膜巨噬细胞中膜褪黑激素受体的特征:百日咳毒素敏感的G蛋白抑制腺苷酸环化酶。

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Melatonin binding sites were characterized in mouse peritoneal macrophages. Binding of 2-[125I]melatonin by macrophages fulfills all criteria for binding to a receptor site. Thus, binding was dependent on time, temperature and cell concentration, stable, reversible, saturable and specific. Stoichiometric studies showed a high-affinity binding site with a Kd of 0.58-0.71 nM. These data are in close agreement with data obtained from kinetic studies (Kd = 0.29 nM). The affinity of these binding sites suggests that they may recognize the physiological concentrations of melatonin in serum. Moreover, binding experiments using macrophage crude membranes showed that melatonin bound specifically to the membranes. Additionally, in competition studies we observed a low-affinity binding site (Kd = 2.02 microM). Melatonin inhibited significantly forskolin-stimulated cyclic AMP accumulation in a dose-dependent manner. This effect was blocked by luzindole, an antagonist of the melatonin membrane receptor. Pretreatment of macrophages with pertussis toxin blocked the inhibitory effect of melatonin. Pertussis toxin ADP-rybosilation and Western blot experiments demonstrated both alpha(i1/2) and alpha(i3/o) G protein subunits expression in mouse peritoneal macrophages membranes. Our results demonstrate the existence of melatonin receptors in mouse peritoneal macrophages, and a pertussis toxin-sensitive melatonin signal transduction pathway that involves the inhibition of adenylyl cyclase.
机译:在小鼠腹膜巨噬细胞中表征了褪黑激素结合位点。巨噬细胞结合2- [125I]褪黑激素符合结合受体位点的所有标准。因此,结合取决于时间,温度和细胞浓度,稳定,可逆,可饱和和特异性。化学计量学研究显示Kd为0.58-0.71 nM的高亲和力结合位点。这些数据与从动力学研究获得的数据(Kd = 0.29 nM)非常吻合。这些结合位点的亲和力表明它们可以识别血清中褪黑激素的生理浓度。此外,使用巨噬细胞粗膜的结合实验表明褪黑激素特异性结合到膜上。此外,在竞争研究中,我们观察到了低亲和力结合位点(Kd = 2.02 microM)。褪黑素以剂量依赖的方式显着抑制毛喉素刺激的环AMP的积累。褪黑激素膜受体的拮抗剂Luzindole阻止了这种作用。百日咳毒素预处理巨噬细胞可阻断褪黑激素的抑制作用。百日咳毒素ADP-rybosilation和Western印迹实验表明,小鼠腹膜巨噬细胞膜中的alpha(i1 / 2)和alpha(i3 / o)G蛋白亚基均表达。我们的结果证明了小鼠腹膜巨噬细胞中褪黑激素受体的存在以及涉及抑制腺苷酸环化酶的百日咳毒素敏感的褪黑素信号转导途径。

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