Phosphoproteomics reveals malaria parasite Protein Kinase G as a signalling hub regulating egress and invasion.
Phosphoproteomics reveals malaria parasite Protein Kinase G as a signalling hub regulating egress and invasion.
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DOI：
10.1038/ncomms8285
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发表时间：
2015-07-07
影响因子：
16.6
通讯作者：
Tobin AB
中科院分区：
文献类型：
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作者：
Alam MM;Solyakov L;Bottrill AR;Flueck C;Siddiqui FA;Singh S;Mistry S;Viskaduraki M;Lee K;Hopp CS;Chitnis CE;Doerig C;Moon RW;Green JL;Holder AA;Baker DA;Tobin AB
我们对人类疟原虫恶性疟原虫中关键的磷酸化依赖性信号通路的了解仍然处于初级阶段。在这里，我们针对重要的 cGMP 依赖性蛋白激酶 PfPKG 解决了这个问题。通过采用化学和遗传工具结合定量全局磷酸化蛋白质组学，我们确定了 69 种蛋白质上的磷酸化位点，这些蛋白质是 PfPKG 的直接或间接细胞靶标。这些 PfPKG 靶标包括参与细胞信号传导、蛋白水解、基因调控、蛋白质输出以及离子和蛋白质运输的蛋白质，表明 cGMP/PfPKG 作为信号传导中枢，在许多核心寄生虫过程中发挥核心作用。我们还表明 PfPKG 活性是寄生虫入侵所必需的。这与钙依赖性蛋白激酶 PfCDPK1 被 PfPKG 磷酸化的发现相关，肌动球蛋白复合物的成分也是如此，为 PfPKG 在寄生虫排出和入侵中的重要作用提供了机制见解。 疟原虫恶性疟原虫表达的蛋白激酶代表了潜在有价值的药物靶点。阿拉姆等人。鉴定其磷酸化依赖于激酶 PfPKG 的蛋白质，揭示了控制寄生虫从红细胞和钙信号传导中流出的调节网络。
Our understanding of the key phosphorylation-dependent signalling pathways in the human malaria parasite, Plasmodium falciparum, remains rudimentary. Here we address this issue for the essential cGMP-dependent protein kinase, PfPKG. By employing chemical and genetic tools in combination with quantitative global phosphoproteomics, we identify the phosphorylation sites on 69 proteins that are direct or indirect cellular targets for PfPKG. These PfPKG targets include proteins involved in cell signalling, proteolysis, gene regulation, protein export and ion and protein transport, indicating that cGMP/PfPKG acts as a signalling hub that plays a central role in a number of core parasite processes. We also show that PfPKG activity is required for parasite invasion. This correlates with the finding that the calcium-dependent protein kinase, PfCDPK1, is phosphorylated by PfPKG, as are components of the actomyosin complex, providing mechanistic insight into the essential role of PfPKG in parasite egress and invasion. Protein kinases expressed by the malaria parasite Plasmodium falciparum represent potentially valuable drug targets. Alam et al. identify proteins whose phosphorylation is dependent on the kinase PfPKG, revealing a regulatory network controlling parasite egress from red blood cells and calcium signalling.