Deep phosphoproteome analysis of Schistosoma mansoni leads development of a kinomic array that highlights sex-biased differences in adult worm protein phosphorylation
Deep phosphoproteome analysis of Schistosoma mansoni leads development of a kinomic array that highlights sex-biased differences in adult worm protein phosphorylation
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DOI：
10.1371/journal.pntd.0008115
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发表时间：
2020-03-01
影响因子：
3.8
通讯作者：
Walker, Anthony J.
中科院分区：
文献类型：
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作者：
Hirst, Natasha L.;Nebel, Jean-Christophe;Walker, Anthony J.
尽管蠕虫寄生虫在世界范围内造成了巨大的痛苦，但我们对蛋白质磷酸化（用于分子信号传导的最重要的翻译后修饰之一）如何调节其稳态和功能知之甚少。这对于抑郁症患者来说尤其如此。在此，我们报告了一个深入的磷酸蛋白质组探索成人曼氏血吸虫，提供了一个最丰富的磷蛋白质资源的任何寄生虫到目前为止，并采用的数据，建立第一个寄生虫特异性激酶组阵列。互补磷酸肽富集策略用于检测映射到3，176个蛋白质的15，844个独特的磷酸肽。磷蛋白被预测参与广泛的生物过程和磷蛋白相互作用组分析揭示了55个高度互连的集群，包括那些富含核糖体，蛋白酶体，吞噬体，剪接体，糖酵解和信号蛋白。鉴定了93个不同的磷酸化基序，其中67个提供了蛋白激酶活性的“足迹”; CaMKII、PKA和CK 1/2高度代表，支持它们对线粒体功能的重要性。在激酶组中，808个磷酸化位点与136个蛋白激酶相匹配，在37个激活环中发现了68个位点。对假定的蛋白激酶-磷蛋白相互作用的分析揭示了典型的网络，但也揭示了信号伴侣之间的新型相互作用。男性和女性成虫提取物的基因组阵列分析显示高磷酸化的转化：转录结构域相关蛋白的两性，和女性的CDK和AMPK肽。此外，八个肽，包括蛋白磷酸酶2C γ，Akt，Rho 2 GT3，SmTK 4，和胰岛素受体更高度磷酸化的女性提取物，强调其可能的重要性，女性蠕虫的功能。我们设想，这些发现，工具和方法将有助于推动新的研究到功能生物学的血吸虫和其他蠕虫寄生虫，并支持努力开发新的治疗方法，为他们的control.Author summarySchistosomes是可怕的寄生虫，导致衰弱和危及生命的疾病人类血吸虫病。我们需要更好地了解这些寄生虫的细胞生物学，以开发新的控制策略。在细胞内，一种称为蛋白质磷酸化的过程控制着分子通讯或“信号”的许多方面，对细胞功能和稳态至关重要。在这里，使用互补的策略，我们已经进行了第一次深入的表征和功能注释的蛋白磷酸化事件的寄生虫，提供了最丰富的磷蛋白资源的任何寄生虫。利用这些知识，我们已经开发出一种新的工具，同时评估这些蠕虫的信号传导过程，并突出性别偏见的差异，成虫蛋白磷酸化。几种蛋白质被发现更大程度上被雌虫提取物磷酸化，这表明它们可能对雌虫功能的重要性。这项工作将有助于推动对寄生虫以及相关寄生虫的基础生物学的新研究，并将支持开发新的药物或疫苗治疗方法以控制它们的努力。
Although helminth parasites cause enormous suffering worldwide we know little of how protein phosphorylation, one of the most important post-translational modifications used for molecular signalling, regulates their homeostasis and function. This is particularly the case for schistosomes. Herein, we report a deep phosphoproteome exploration of adult Schistosoma mansoni, providing one of the richest phosphoprotein resources for any parasite so far, and employ the data to build the first parasite-specific kinomic array. Complementary phosphopeptide enrichment strategies were used to detect 15,844 unique phosphopeptides mapping to 3,176 proteins. The phosphoproteins were predicted to be involved in a wide range of biological processes and phosphoprotein interactome analysis revealed 55 highly interconnected clusters including those enriched with ribosome, proteasome, phagosome, spliceosome, glycolysis, and signalling proteins. 93 distinct phosphorylation motifs were identified, with 67 providing a 'footprint' of protein kinase activity; CaMKII, PKA and CK1/2 were highly represented supporting their central importance to schistosome function. Within the kinome, 808 phosphorylation sites were matched to 136 protein kinases, and 68 sites within 37 activation loops were discovered. Analysis of putative protein kinase-phosphoprotein interactions revealed canonical networks but also novel interactions between signalling partners. Kinomic array analysis of male and female adult worm extracts revealed high phosphorylation of transformation:transcription domain associated protein by both sexes, and CDK and AMPK peptides by females. Moreover, eight peptides including protein phosphatase 2C gamma, Akt, Rho2 GTPase, SmTK4, and the insulin receptor were more highly phosphorylated by female extracts, highlighting their possible importance to female worm function. We envision that these findings, tools and methodology will help drive new research into the functional biology of schistosomes and other helminth parasites, and support efforts to develop new therapeutics for their control.Author summarySchistosomes are formidable parasites that cause the debilitating and life-threatening disease human schistosomiasis. We need to better understand the cellular biology of these parasites to develop novel strategies for their control. Within cells, a process called protein phosphorylation controls many aspects of molecular communication or 'signalling' and is central to cellular function and homeostasis. Here, using complementary strategies, we have performed the first in-depth characterisation and functional annotation of protein phosphorylation events in schistosomes, providing one of the richest phosphoprotein resources for any parasite to date. Using this knowledge, we have developed a novel tool to simultaneously evaluate signalling processes in these worms and highlight sex-biased differences in adult worm protein phosphorylation. Several proteins were found to be more greatly phosphorylated by female worm extracts, suggesting their possible importance to female worm function. This work will help drive new research into the fundamental biology of schistosomes, as well as related parasites, and will support efforts to develop new drug or vaccine-based therapeutics for their control.