Structural basis for the interaction of diapause hormone with its receptor in the silkworm, Bombyx mori
Structural basis for the interaction of diapause hormone with its receptor in the silkworm, Bombyx mori
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家蚕滞育激素与其受体相互作用的结构基础
DOI：
10.1096/fj.201700931r
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发表时间：
2018-03-01
期刊：
影响因子：
4.8
通讯作者：
Zhou，Naiming
中科院分区：
文献类型：
--
作者：
Shen，Zhangfei;Jiang，Xue;Zhou，Naiming
滞育激素 (DH) 是一种 24-aa 酰胺化神经肽，通过与其受体 BommoDH 受体 (Bommo-DHR) 敏感且选择性的相互作用，引发家蚕 (Bombyx mori) (Bommo) 胚胎滞育。以往对Bommo-DH构效关系的研究都是基于体内滞育诱导生物测定，对Bommo-DHR的结构或其与DH的迭代提供的信息很少。在这里，为了揭示 Bommo-DH 与其受体的相互作用，化学合成了 Bommo-DH 的 N 端截短类似物和丙氨酸扫描突变体，并使用 Cy5.5 标记的 Bommo-DH 竞争性结合测定和基于 Bommo-DHR 的功能测定（包括 cAMP 测定和 Ca2+ 动员测定）进行功能评估。我们的研究表明，Bommo-DH 的 C 端残基 Arg23 和 Leu24 对于 Bommo-DHR 的结合和激活至关重要，Trp19 和 Phe20 也有助于 Bommo-DH 的功能活性。相反，当 Gly21 或 Pro22 被丙氨酸取代时，两种突变体都表现出与野生型肽无法区分的结合和信号传导活性。此外，我们的同源建模和分子动力学模拟以及实验验证已经确定了 Bommo-DHR 中与 Bommo-DH 相互作用至关重要的 Glu89、Phe172、Phe194 和 Tyr299 残基。这些结果可能加深我们对A类GPCR与其肽配体相互作用的理解，特别是信息素生物合成激活神经肽/DH家族神经肽与其同源受体之间的相互作用。-Shen, Z., Jiang, X., Yan, L., Chen, Y., Wang, W., Shi, Y., Shi, L., Liu, D., Zhou, N. 滞育相互作用的结构基础激素及其在家蚕（Bombyx mori）中的受体。 FASEB J. 32, 1338-1353 (2018)。 www.fasebj.org
Diapause hormone (DH) is a 24‐aa amidated neuropeptide that elicits the embryonic diapause of the silkworm,Bombyx mori (Bommo), viasensitive and selective interaction with its receptor,BommoDH receptor (Bommo‐DHR). Previous studies of the structure‐activity relationship ofBommo‐DH were all based on anin vivodiapause‐induction bioassay, which has provided little information on the structure ofBommo‐DHR or its iteration with DH. Here, to unveil the interaction ofBommo‐DH with its receptor, N‐terminally truncated analogs and alanine‐scanning mutants ofBommo‐DH were chemically synthesized and functionally evaluated by using a Cy5.5‐ labeledBommo‐DH competitive binding assay andBommo‐DHR‐based functional assays, including cAMP assay and Ca2+mobilization assay. Our study demonstrates that the C‐terminal residues of Arg23 and Leu24 ofBommo‐DH are essential for the binding and activation ofBommo‐DHR, and that Trp19 and Phe20 also contribute to the functional activity ofBommo‐DH. In contrast, when Gly21 or Pro22 were replaced with alanine, both mutants exhibited binding and signaling activities that were indistinguishable from the wild‐type peptide. Furthermore, our homology modeling and molecular dynamics simulations, together with experimental validations, have identified the residues of Glu89, Phe172, Phe194, and Tyr299 inBommo‐DHR that are critically involved in the interaction withBommo‐DH. These results may deepen our understanding of the interactions of class‐A GPCRs with their peptidic ligands, particularly those between pheromone biosynthesis‐activating neuropeptide/DH family neuropeptides and their cognate receptors.—Shen, Z., Jiang, X., Yan, L., Chen, Y., Wang, W., Shi, Y., Shi, L., Liu, D., Zhou, N. Structural basis for the interaction of diapause hormone with its receptor in the silkworm, Bombyx mori. FASEB J. 32, 1338‐1353 (2018). www.fasebj.org