Department of Zoology, University of Delhi, Delhi, India.
PLoS One. 2022 Jul 7;17(7):e0270881. doi: 10.1371/journal.pone.0270881. eCollection 2022.
The present study, in addition to molecular characterization of leptin (lepa) and its receptor (lepr) of spotted snakehead Channa punctata, is focussed on physicochemical, structural, evolutionary and selection pressure analyses which are poorly elucidated in teleosts in spite of that existence of these genes is well reported in several fish species. The putative full-length Lep and Lepr of C. punctata showed conserved structural and functional domains, especially the residues responsible for structural integrity and signal transduction. Conversely, residues predicted essential for Lep-Lepr interaction displayed divergence between teleosts and tetrapods. Impact of substitutions/deletions predicted using protein variation effect analyser tool highlighted species specificity in ligand-receptor interaction. Physicochemical properties of ligand and receptor predicted for the first time in vertebrates revealed high aliphatic and instability indices for both Lepa and Lepr, indicating thermostability of proteins but their instability under ex vivo conditions. Positive grand average of hydropathy score of Lepa suggests its hydrophobic nature conjecturing existence of leptin binding proteins in C. punctata. In addition to disulphide bonding, a novel posttranslational modification (S-126 phosphorylation) was predicted in Lepa of C. punctata. In Lepr, disulphide bond formation and N-linked glycosylation near WSXWS motif in ECD, and phosphorylation at tyrosine residues in ICD were predicted. Leptin and its receptor sequence of C. punctata cladded with its homolog from C. striata and C. argus of order Anabantiformes. Leptin system of Anabantiformes was phylogenetically closer to that of Pleuronectiformes, Scombriformes and Perciformes. Selection pressure analysis showed higher incidence of negative selection in teleostean leptin genes indicating limited adaptation in their structure and function. However, evidence of pervasive and episodic diversifying selection laid a foundation of co-evolution of Lepa and Lepr in teleosts.
本研究除了对斑点叉尾鮰瘦素(lepa)及其受体(lepr)进行分子特征描述外,还集中于对理化性质、结构、进化和选择压力进行分析,尽管这些基因在几种鱼类中已有报道,但在硬骨鱼类中这些方面的研究仍不够深入。斑点叉尾鮰的推定全长 Lep 和 Lepr 具有保守的结构和功能域,特别是负责结构完整性和信号转导的残基。相反,预测对 Lep-Lepr 相互作用至关重要的残基在硬骨鱼类和四足动物之间存在差异。使用蛋白质变异效应分析工具预测的取代/缺失的影响突出了配体-受体相互作用的物种特异性。首次在脊椎动物中预测配体和受体的理化性质表明,Lepa 和 Lepr 的亲脂性和不稳定性指数都很高,这表明蛋白质的热稳定性,但它们在体外条件下不稳定。Lepa 的平均亲水性评分呈正值,表明其疏水性,推测斑点叉尾鮰中存在瘦素结合蛋白。除了二硫键外,还预测到斑点叉尾鮰 Lepa 中的一种新的翻译后修饰(S-126 磷酸化)。在 Lepr 中,预测到 ECD 中 WSXWS 基序附近形成二硫键和 N-连接糖基化,以及 ICD 中酪氨酸残基磷酸化。斑点叉尾鮰的瘦素及其受体序列与斑叉尾鮰和尖嘴鲈的同源序列聚类,它们同属于鲈形目。鲈形目的瘦素系统在系统发育上与鲽形目、鲭形目和鲈形目更为接近。选择压力分析表明,硬骨鱼类瘦素基因的负选择发生率更高,表明其结构和功能的适应性有限。然而,普遍存在的和偶发的多样化选择的证据为 Lepa 和 Lepr 在硬骨鱼类中的共同进化奠定了基础。