Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea.
Molecular cloning and functional characterization of the diapause hormone receptor in the corn earworm Helicoverpa zea.
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DOI：
10.1016/j.peptides.2013.11.005
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发表时间：
2014-03
期刊：
影响因子：
3
通讯作者：
Park Y
中科院分区：
文献类型：
--
作者：
Jiang H;Wei Z;Nachman RJ;Park Y
家蚕蛾体内的滞育激素(Dh)已显示出终止蛹滞育的活性，而家蚕中的滞育激素(Dh)具有诱导胚胎滞育的作用。在本研究中，我们从玉米穗虫Helicoverpa zea(HzDHr)中克隆了滞育激素受体(HzDHr)，并在异源报告系统中测试了其配体特异性。HzDHr在中国仓鼠卵巢(CHO)细胞中表达，并与Aequorin报告基因共转染CHO细胞，用于检测其配体活性。共有68种化学物质，包括天然的水解酶类似物和结构相似的多肽模拟物，被测试激动剂和拮抗剂的活性。用2-氨基-7-溴基-琥珀酰基(2Abf-Suc)N端修饰的几种多肽模拟物显示出很强的激动活性，包括2Abf-Suc-F[da]PRLamide、2Abf-Suc-F[DR]PRlamide、2Abf-Suc-FKPRlamide和2Abf-Suc-FGPRlamide。黑腹果蝇蜕皮触发激素(FFLKITKNVPRLamide)具有拮抗活性。有趣的是，HzDHr并不区分DH(WFGPRLamide C-末端基序)和另一种密切相关的内源性多肽-焦激肽1(FXPRXide；与WFGPRLamide分开的C-末端基序)。我们提供了大规模的体外数据，作为开发激动剂和拮抗剂来扰乱水解酶信号通路的参考。
The diapause hormone (DH) in the heliothine moth has shown its activity in termination of pupal diapause, while the orthology in the silkworm is known to induce embryonic diapause. In the current study, we cloned the diapause hormone receptor from the corn earworm Helicoverpa zea (HzDHr) and tested its ligand specificities in a heterologous reporter system. HzDHr was expressed in Chinese Hamster Ovary (CHO) cells, which were co-transfected with the aequorin reporter, and was used to measure the ligand activities. A total of 68 chemicals, including natural DH analogs and structurally similar peptide mimetics, were tested for agonistic and antagonistic activities. Several peptide mimetics with a 2-amino-7-bromofluorene-succinoyl (2Abf-Suc) N-terminal modification showed strong agonistic activities; these mimetics included 2Abf-Suc-F[dA]PRLamide, 2Abf-Suc-F[dR]PRLamide, 2Abf-Suc-FKPRLamide and 2Abf-Suc-FGPRLamide. Antagonistic activity was found in the ecdysis triggering hormone in Drosophila melanogaster (FFLKITKNVPRLamide). Interestingly, HzDHr does not discriminate between DH (WFGPRLamide C-terminal motif) and another closely related endogenous peptide, pyrokinin 1 (FXPRXamide; a C-terminal motif that is separate from WFGPRLamide). We provide large-scale in vitro data that serve as a reference for the development of agonists and antagonists to disrupt the DH signaling pathway.
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