Removal of partial agonism from parathyroid hormone (PTH)-related protein-(7-34)NH2 by substitution of PTH amino acids at positions 10 and 11.
Removal of partial agonism from parathyroid hormone (PTH)-related protein-(7-34)NH2 by substitution of PTH amino acids at positions 10 and 11.
复制标题
通过替换第 10 和 11 位 PTH 氨基酸，去除甲状旁腺激素 (PTH) 相关蛋白 -(7-34)NH2 的部分激动作用。
DOI：
10.1210/endo-127-1-491
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发表时间：
1990
期刊：
影响因子：
4.8
通讯作者：
R. Mckee
中科院分区：
文献类型：
--
作者：
R. Nutt;M. Caulfield;J. Levy;S. W. Gibbons;M. Rosenblatt;R. Mckee
PTHrP(7-34)NH2和[D-Trp12]PTHrP(7-34)NH2已被证明是比相应的PTH多肽[Tyr34]bpth(7-34)NH2更有效的拮抗剂。然而，这些多肽在ROS 17/2.8细胞中也表现出对腺苷环化酶活性的部分兴奋作用。在这项研究中，通过从PTHrP(7-34)NH2中去除激动剂，在保留拮抗剂效力的情况下，设计了一种纯的有效的PTH和PTHrP拮抗剂。由于[Tyr34]bpth(7-34)NH2缺乏激动剂活性，我们将PTH序列中的两个天然氨基酸引入到PTHrP和强大的D-Trp12类似物中各自的位置。在ROS细胞中，[Asn10Leu11]-和[Asn10，leu11，D-Trp12]-PTHrP(7-34)NH2的拮抗剂作用分别是PTHrP(7-34)NH2和[D-Trp12]PTHrP(7-34)NH2的23倍和26倍。此外，即使在用地塞米松和百日咳毒素处理的高反应性细胞上进行的检测中，这些多肽也没有表现出部分激活性。相反，当PTHrP序列Asp10，Lys11插入到[Tyr34]hPTH(7-34)NH2中时，拮抗剂的效力下降了6倍以上，并获得了类似PTH的激动剂活性。这些结果表明，甲状旁腺激素和甲状旁腺素rP的激活结构域都可以扩展到包括1-12区，并且10-12区除了N-末端六肽外，不仅对受体结合重要，而且对激素信号转导也很重要。
PTHrP(7-34)NH2 and [D-Trp12]PTHrP(7-34)NH2 have previously been shown to be shown to be more potent antagonists than the corresponding PTH peptide, [Tyr34]bPTH(7-34)NH2. However, these peptides also display partial agonism for adenylate cyclase activity in ROS 17/2.8 cells. In this study, design of a pure potent antagonist of PTH and PTHrP by removal of agonism from PTHrP(7-34)NH2 with retention of antagonist potency was accomplished. Since [Tyr34]bPTH(7-34)NH2 lacks agonist activity, we introduced two amino acids native to the PTH sequence into their respective positions in PTHrP and the potent D-Trp12 analog. [Asn10Leu11]- and [Asn10,leu11,D-Trp12]-PTHrP(7-34)NH2 were found to be 23- and 26-fold more potent as antagonists in ROS cells than PTHrP(7-34)NH2 and [D-Trp12]PTHrP(7-34)NH2, respectively. In addition, these peptides did not display partial agonism, even in an assay based on highly responsive cells pretreated with dexamethasone and pertussis toxin. In contrast, when the PTHrP sequence Asp10,Lys11 was inserted into [Tyr34]hPTH(7-34)NH2, antagonist potency declined by more than 6-fold and PTH-like agonist activity was installed. These results demonstrate that the activation domain of both PTH and PTHrP can be extended to include the 1-12 region and that the 10-12 region, in addition to the N-terminal hexapeptide, is important not only for receptor binding but also for hormonal signal transduction.