Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents.
Rickettsia prowazekii methionine aminopeptidase as a promising target for the development of antibacterial agents.
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DOI：
10.1016/j.bmc.2016.11.013
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发表时间：
2017-02-01
影响因子：
3.5
通讯作者：
Hagen TJ
中科院分区：
文献类型：
--
作者：
Helgren TR;Chen C;Wangtrakuldee P;Edwards TE;Staker BL;Abendroth J;Sankaran B;Housley NA;Myler PJ;Audia JP;Horn JR;Hagen TJ
甲氨酰氨基肽酶（MetAP）是一类普遍存在的酶，对于许多细菌物种的生存至关重要。这些酶负责从新生蛋白质切割N-末端甲酰基-甲硫氨酸起始物，以引发通常对适当蛋白质功能至关重要的翻译后修饰。因此，抑制MetAP活性已被认为是一种新的抗菌靶标。我们在本研究中通过靶向专性细胞内病原体普氏立克次体中的MetAP酶来测试这一想法。我们首先通过采用体外酶活性测定鉴定了有效的RpMetAP抑制物质。然后利用分子对接程序AutoDock将已发表的抑制的MetAP物质的晶体结构与RpMetAP的对接位姿进行比较。基于这些计算机模拟和体外筛选，测试了23种化合物的子集对R.在肺血管内皮细胞（EC）培养物感染模型系统中的prowazekii生长。在确定对EC无毒的浓度范围内测试所有化合物，并且23种化合物中的10种显示出对R的显著抑制。prowazekii生长这些数据突出了抑制RpMetAP作为一种新型抗菌策略的治疗潜力，并为临床前感染动物模型的未来研究奠定了基础。
Methionine aminopeptidase (MetAP) is a class of ubiquitous enzymes essential for the survival of numerous bacterial species. These enzymes are responsible for the cleavage of N-terminal formyl-methionine initiators from nascent proteins to initiate post-translational modifications that are often essential to proper protein function. Thus, inhibition of MetAP activity has been implicated as a novel antibacterial target. We tested this idea in the present study by targeting the MetAP enzyme in the obligate intracellular pathogen Rickettsia prowazekii. We first identified potent RpMetAP inhibitory species by employing an in vitro enzymatic activity assay. The molecular docking program AutoDock was then utilized to compare published crystal structures of inhibited MetAP species to docked poses of RpMetAP. Based on these in silico and in vitro screens, a subset of 23 compounds was tested for inhibition of R. prowazekii growth in a pulmonary vascular endothelial cell (EC) culture infection model system. All compounds were tested over concentration ranges that were determined to be non-toxic to the ECs and 10 of the 23 compounds displayed substantial inhibition of R. prowazekii growth. These data highlight the therapeutic potential for inhibiting RpMetAP as a novel antimicrobial strategy and set the stage for future studies in pre-clinical animal models of infection.