SAM domain-dependent activity of PfTKL3, an essential tyrosine kinase-like kinase of the human malaria parasite Plasmodium falciparum
SAM domain-dependent activity of PfTKL3, an essential tyrosine kinase-like kinase of the human malaria parasite Plasmodium falciparum
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DOI：
10.1007/s00018-010-0434-3
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发表时间：
2010-10-01
影响因子：
8
通讯作者：
Doerig, Christian
中科院分区：
文献类型：
--
作者：
Abdi, Abdirahman;Eschenlauer, Sylvain;Doerig, Christian
在过去的十年中，几种蛋白激酶抑制剂已经进入癌症化疗市场。病原体的激酶组是传染病中潜在的有吸引力的靶标。恶性疟原虫是一种寄生原生生物，是人类疟疾的最致命形式，其大多数蛋白激酶的功能仍不清楚。在这里，我们提出了一个彻底的表征PfTKL 3（PF13_0258），一种酶，属于酪氨酸激酶样激酶（TKL）组。我们通过反向遗传学证明PfTKL 3对人类红细胞中的无性寄生虫增殖至关重要。PfTKL 3在无性和配子体阶段都有表达，在配子体阶段，蛋白质与细胞骨架微管共定位。重组PfTKL 3显示体外自磷酸化活性，并能够磷酸化外源性底物，这两种活性都显著依赖于N-末端“无菌α-基序”结构域的存在。这项研究确定PfTKL 3作为一个经过验证的药物靶点，适合高通量筛选。
Over the last decade, several protein kinases inhibitors have reached the market for cancer chemotherapy. The kinomes of pathogens represent potentially attractive targets in infectious diseases. The functions of the majority of protein kinases of Plasmodium falciparum, the parasitic protist responsible for the most virulent form of human malaria, remain unknown. Here we present a thorough characterisation of PfTKL3 (PF13_0258), an enzyme that belongs to the tyrosine kinase-like kinase (TKL) group. We demonstrate by reverse genetics that PfTKL3 is essential for asexual parasite proliferation in human erythrocytes. PfTKL3 is expressed in both asexual and gametocytes stages, and in the latter the protein co-localises with cytoskeleton microtubules. Recombinant PfTKL3 displays in vitro autophosphorylation activity and is able to phosphorylate exogenous substrates, and both activities are dramatically dependent on the presence of an N-terminal "sterile alpha-motif" domain. This study identifies PfTKL3 as a validated drug target amenable to high-throughput screening.