Achieving signalling selectivity of ligands for the corticotropin-releasing factor type 1 receptor by modifying the agonist's signalling domain
Achieving signalling selectivity of ligands for the corticotropin-releasing factor type 1 receptor by modifying the agonist's signalling domain
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DOI：
10.1038/sj.bjp.0707293
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发表时间：
2007-07-01
影响因子：
7.3
通讯作者：
Berger, H.
中科院分区：
文献类型：
--
作者：
Beyermann, M.;Heinrich, N.;Berger, H.
背景和目的：大多数药物靶向G蛋白偶联受体（GPCR），其通常可以激活不同的信号传导事件。本研究的目的是实现促肾上腺皮质激素释放因子受体1型（CRF 1）配体的功能选择性。我们系统地取代尿皮质素，一种天然的肽激动剂的CRF 1，与庞大的氨基酸，（苯甲酰基-苯丙氨酸，萘基丙氨酸），并使用受体结合测定类似物对人胚肾细胞中CRF 1与GS-和G（i）-蛋白偶联的作用，[S-35]-GTPgS结合刺激和cAMP积累测定。关键结果：天然配体通过CRF 1刺激G（s）和G（i）活化，导致cAMP积累的刺激和抑制。在尿皮质素的第6 - 15位上的单取代，依赖于取代基的位置和性质，导致保留G（s）活性的配体，但没有G（i）活性，仅刺激cAMP积累，并竞争性地拮抗sauvagine对G（i）的激活。与此相反，类似物与此序列以外的取代非选择性激活G（S）和G（i），urocortin does.Conclusions和影响：修改在一个特定的区域，我们称之为信号结构域，在多肽激动剂urocortin导致类似物，表现为激动剂，并在同一时间，拮抗剂不同的G-蛋白的激活CRF 1。这一发现意味着当与不同的G蛋白偶联时，受体的活性构象之间存在显着差异。在其他多肽激素受体配体中的信号传导信息的类似结构编码将导致用于开发信号传导选择性候选药物的一般概念。
Background and purpose: Most of the pharmaceuticals target G-protein-coupled receptors (GPCRs) which can generally activate different signalling events. The aim of this study was to achieve functional selectivity of corticotropin-releasing factor receptor type 1 (CRF1) ligands.Experimental approach: We systematically substituted urocortin, a natural peptide agonist of CRF1, with bulky amino acids (benzoyl-phenylalanine, naphthylalanine) and determined the effect of the analogues on coupling of CRF1 to Gs-and G(i)-protein in human embryonic kidney cells, using receptor binding, [S-35]-GTPgS binding stimulation, and cAMP accumulation assays.Key results: Native ligands stimulated G(s) and G(i) activation through CRF1, resulting in stimulation and then inhibition of cAMP accumulation. Single replacements in urocortin at positions 6 -15 led, dependent on the position and nature of the substituent, to ligands that conserved G(s) activity, but were devoid of G(i) activity, only stimulating cAMP accumulation, and competitively antagonized the G(i) activation by sauvagine. In contrast, analogues with substitutions outside this sequence non-selectively activated G(s) and G(i), as urocortin did.Conclusions and implications: Modifications in a specific region, which we have called the signalling domain, in the polypeptide agonist urocortin resulted in analogues that behaved as agonists and, at the same time, antagonists for the activation of different G-proteins by CRF1. This finding implies significant differences between active conformations of the receptor when coupled to different G-proteins. A similar structural encoding of signalling information in other polypeptide hormone receptor ligands would result in a general concept for the development of signalling-selective drug candidates.