Helgren Travis R, Chen Congling, Wangtrakuldee Phumvadee, Edwards Thomas E, Staker Bart L, Abendroth Jan, Sankaran Banumathi, Housley Nicole A, Myler Peter J, Audia Jonathon P, Horn James R, Hagen Timothy J
Department of Chemistry and Biochemistry, Northern Illinois University, 1425 W. Lincoln Hwy, DeKalb, IL 60115, USA.
Beryllium Discovery Corp., 7869 NE Day Road West, Bainbridge Island, WA 98110, USA; Seattle Structural Genomics Center for Infectious Disease (SSGCID), Seattle, WA, USA.
Bioorg Med Chem. 2017 Feb 1;25(3):813-824. doi: 10.1016/j.bmc.2016.11.013. Epub 2016 Nov 10.
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 17 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 8 of the 17 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.
甲硫氨酸氨肽酶(MetAP)是一类广泛存在的酶,对众多细菌物种的生存至关重要。这些酶负责从新生蛋白质中切割N端甲酰甲硫氨酸起始子,以启动翻译后修饰,而这些修饰通常对蛋白质的正常功能至关重要。因此,抑制MetAP活性已被认为是一种新型抗菌靶点。在本研究中,我们通过靶向专性细胞内病原体普氏立克次体中的MetAP酶来验证这一想法。我们首先通过体外酶活性测定法鉴定出强效的RpMetAP抑制物种。然后利用分子对接程序AutoDock将已发表的被抑制的MetAP物种的晶体结构与RpMetAP的对接姿势进行比较。基于这些计算机模拟和体外筛选,在肺血管内皮细胞(EC)培养感染模型系统中测试了17种化合物的一个子集对普氏立克次体生长的抑制作用。所有化合物都在确定对EC无毒的浓度范围内进行了测试,17种化合物中有8种对普氏立克次体生长表现出显著抑制作用。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,并为未来在临床前感染动物模型中的研究奠定了基础。