鲤鱼促性腺激素抑制激素受体（GnIHR2）的鉴定、对接和分子动力学模拟。
Characterization, Docking and Molecular Dynamics Simulation of Gonadotropin-Inhibitory Hormone Receptor (GnIHR2) in Labeo Catla.
机构信息
Department of Aquaculture, College of Fisheries, Dr. Rajendra Prasad Central Agricultural University, Pusa, Bihar, India.
ICAR-Central Institute of Fisheries Education, Mumbai, India.
出版信息
Cell Physiol Biochem. 2020 Sep 2;54(5):825-841. doi: 10.33594/000000272.
BACKGROUND/AIMS: GnIH receptors (GnIHRs) belong to the family of G-protein coupled receptors (GPCRs) and play a key role in the regulation of reproduction from fishes to mammals, either by inhibiting or stimulating the expression of gonadotropins. The aim of this study was to characterize GnIH receptor (GnIHR2) from Indian Major Carp, Labeo catla and its docking and simulation with GnIH antagonist RF313.
METHODS
The full length sequence of GnIHR2 was obtained with RACE PCR. The docking analysis of RF313 with GnIHR2 receptor was performed with AutoDock v. 4.2.6 and molecular dynamics (MD) simulation with GROMACS 5.0.
RESULTS
In the present study, we cloned full-length cDNA (1733 bp) of GnIHR2 from the brain of L. catla. The phylogenetic analysis showed clustering of catla GnIHR2 with goldfish and zebrafish in the GPR147 group. L. catla GnIHR2 receptor comprised seven transmembrane domains and the 3D-structure was predicted by I-TASSER tool. The docking analysis revealed high binding affinity (-11.6 kcal/mol) of GnIH antagonist, RF313 towards GnIHR2 receptor. The primary bonds involved were alkyl and hydrogen bonds while the amino acids participated were proline 43, 210, 339, cysteine 214, leucine 211, serine 213 and phenylalanine 338. The MD simulation analysis of docked complex for 100 nano-seconds (ns) in the lipid membrane environment showed the stability of the complex with time.
CONCLUSION
Our study showed that GnIH antagonist, RF313 interact tightly with the GnIH receptor, GnIHR2 of L. catla. To the best of our knowledge, this is the first report on computational modelling and MD simulation of GnIH receptor in fishes. This will help in functional characterization studies of GnIH/GnIHR system in vertebrates.
背景/目的：GnIH 受体（GnIHRs）属于 G 蛋白偶联受体（GPCRs）家族，在从鱼类到哺乳动物的生殖调控中发挥着关键作用，通过抑制或刺激促性腺激素的表达。本研究的目的是从印度鲤鱼（Labeo catla）中鉴定 GnIH 受体（GnIHR2），并对其与 GnIH 拮抗剂 RF313 的对接和模拟。
方法
采用 RACE PCR 获得 GnIHR2 的全长序列。采用 AutoDock v. 4.2.6 进行 RF313 与 GnIHR2 受体的对接分析，采用 GROMACS 5.0 进行分子动力学（MD）模拟。
结果
本研究从 L. catla 脑中克隆出 GnIHR2 的全长 cDNA（1733bp）。系统发育分析表明，catla GnIHR2 与金鱼和斑马鱼聚在 GPR147 组中。L. catla GnIHR2 受体包含七个跨膜域，采用 I-TASSER 工具预测其 3D 结构。对接分析显示 GnIH 拮抗剂 RF313 对 GnIHR2 受体具有高结合亲和力（-11.6 kcal/mol）。涉及的主要键为烷基和氢键，参与的氨基酸为脯氨酸 43、210、339、半胱氨酸 214、亮氨酸 211、丝氨酸 213 和苯丙氨酸 338。在脂质膜环境中对对接复合物进行 100 纳秒（ns）的 MD 模拟分析表明，复合物随时间稳定。
结论
我们的研究表明，GnIH 拮抗剂 RF313 与 L. catla 的 GnIH 受体 GnIHR2 紧密相互作用。据我们所知，这是鱼类 GnIH 受体的计算建模和 MD 模拟的首次报道。这将有助于研究脊椎动物中 GnIH/GnIHR 系统的功能特征。
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