Functional differences between NPFF1 and NPFF2 receptor coupling:: High intrinsic activities of RFamide-related peptides on stimulation of [35S]GTPγS binding
Functional differences between NPFF1 and NPFF2 receptor coupling:: High intrinsic activities of RFamide-related peptides on stimulation of [35S]GTPγS binding
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DOI：
10.1016/j.neuropharm.2006.07.034
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发表时间：
2007-02-01
影响因子：
4.7
通讯作者：
Zajac, J. -M.
中科院分区：
文献类型：
--
作者：
Gouarderes, C.;Mazarguil, H.;Zajac, J. -M.
通过优化的[S-35]GTP γ S结合试验，研究了属于RF酰胺家族的三个成员的肽：神经肽FF（NPFF）、催乳素释放肽（PrRP）和26 RF酰胺对在中国人卵巢（CHO）细胞中稳定表达的NPFF 1和NPFF 2受体的功能活性（效力和功效）。尽管它们在亲和力和选择性方面存在很大差异，但所有测试的类似物都表现为对NPFF 1和NPFF 2受体的激动剂。在测定中高NaCl浓度强烈地增加了对NPFF 2受体的功效，并增加了激动剂之间的差异。在低钠条件下，尽管激动剂的效力与它们对NPFF 1受体的亲和力相关，但NPFF 2受体表现出非凡的活性，因为所有测试的化合物显示出GTP γ S结合的EC 50值低于它们的Ki值。NPFF 1和NPFF受体之间的功能值的比较揭示了意想不到的有效的选择性NPFF 2激动剂，特别是对于PLRF酰胺和VGRF酰胺序列。通过使用阻断肽，我们还表明，GOCD和Gas是NPFF 1受体的主要转换器，而NPFF 2可能与G α（i2），G α（i3），G α（o）和G α（s）蛋白偶联。我们的数据表明，NPPF 1和NPFF 2受体是不同的耦合到G蛋白在CHO细胞。(c)2006爱思唯尔有限公司保留所有权利。
By using an optimized [S-35]GTP gamma S binding assay, the functional activities (potency and efficacy) of peptides belonging to three members of the RFamide family; Neuropeptide FF (NPFF), prolactin-releasing peptide (PrRP) and 26RFamide, were investigated on NPFF1 and NPFF2 receptors stably expressed in Chinese Hamster Ovary (CHO) cells. Despite their large differences in affinity and selectivity, all analogues tested behaved as agonists toward NPFF1 and NPFF2 receptors. High NaCl concentration in the assay strongly increased the efficacy toward NPFF2 receptors and augmented differences among agonists. In low sodium conditions, whereas the potencies of agonists correlated with their affinities for NPFF1 receptors, NPFF2 receptors exhibited an extraordinary activity since all compounds tested displayed EC50 values of GTP gamma S binding lower than their K, values. Comparisons of functional values between NPFF1 and NPFF, receptors revealed unexpected potent selective NPFF2 agonists especially for the PLRFamide and the VGRFamide sequences. By using blocker peptides, we also show that GOCD and Gas are the main transducers of NPFF1 receptors while NPFF2 are probably coupled with G alpha(i2), G alpha(i3), G alpha(o) and G alpha(s) proteins. Our data indicate that NPPF1 and NPFF2 receptors are differently coupled to G proteins in CHO cells. (c) 2006 Elsevier Ltd. All rights reserved.