Roth Adelheid L, Marzola Erika, Rizzi Anna, Arduin Marika, Trapella Claudio, Corti Corrado, Vergura Raffaella, Martinelli Prisca, Salvadori Severo, Regoli Domenico, Corsi Mauro, Cavanni Paolo, Caló Girolamo, Guerrini Remo
Department of Biology, Psychiatry Center of Excellence for Drug Discovery, GlaxoSmithKline Pharmaceuticals, Via Alessandro Fleming 2, 37135 Verona, Italy.
Department of Pharmaceutical Science and Biotechnology Center, University of Ferrara, Via Fossato di Mortara 19, 44100 Ferrara, Italy.
J Biol Chem. 2006 Jul 28;281(30):20809-20816. doi: 10.1074/jbc.M601846200. Epub 2006 May 23.
Neuropeptide S (NPS) has been recently recognized as the endogenous ligand for the previous orphan G-protein-coupled receptor GPR154, now referred to as the NPS receptor (NPSR). The NPS-NPSR receptor system regulates important biological functions such as sleeping/wakening, locomotion, anxiety, and food intake. To collect information on the mechanisms of interaction between NPS and its receptor, a classical structure-activity relationship study was performed. Human (h) NPS derivatives obtained by Ala and d-scan and N- and C-terminal truncation were assessed for their ability to stimulate calcium release in HEK293 cells expressing the human recombinant NPSR. The results of this study indicate that (i) the effect of hNPS is mimicked by the fragment hNPS-(1-10); (ii) Phe(2), Arg(3), and Asn(4) are crucial for biological activity; (iii) the sequence Thr(8)-Gly(9)-Met(10) is important for receptor activation, although with non-stringent chemical requirements; and (iv) the sequence Val(6)-Gly(7) acts as a hinge region between the two above-mentioned domains. However, the stimulatory effect of hNPS given intracerebroventricularly on mouse locomotor activity was not fully mimicked by hNPS-(1-10), suggesting that the C-terminal region of the peptide maintains importance for in vivo activity. In conclusion, this study identified the amino acid residues of this peptide most important for receptor activation.
神经肽S（NPS）最近被确认为先前孤儿G蛋白偶联受体GPR154（现称为NPS受体，NPSR）的内源性配体。NPS-NPSR受体系统调节重要的生物学功能，如睡眠/觉醒、运动、焦虑和食物摄入。为了收集有关NPS与其受体相互作用机制的信息，进行了经典的构效关系研究。通过丙氨酸扫描和d扫描以及N端和C端截短获得的人（h）NPS衍生物，在表达人重组NPSR的HEK293细胞中评估其刺激钙释放的能力。这项研究的结果表明：（i）hNPS的作用可被片段hNPS-(1-10)模拟；（ii）苯丙氨酸（2）、精氨酸（3）和天冬酰胺（4）对生物活性至关重要；（iii）序列苏氨酸（8）-甘氨酸（9）-甲硫氨酸（10）对受体激活很重要，尽管化学要求不严格；（iv）序列缬氨酸（6）-甘氨酸（7）作为上述两个结构域之间的铰链区。然而，脑室内给予hNPS对小鼠运动活性的刺激作用并未被hNPS-(1-10)完全模拟，这表明该肽的C端区域对体内活性仍然很重要。总之，这项研究确定了该肽中对受体激活最重要的氨基酸残基。
