Olaleye Omonike, Raghunand Tirumalai R, Bhat Shridhar, He Jian, Tyagi Sandeep, Lamichhane Gyanu, Gu Peihua, Zhou Jiangbing, Zhang Ying, Grosset Jacques, Bishai William R, Liu Jun O
Department of Pharmacology and Molecular Sciences, Johns Hopkins School of Medicine, Baltimore, MD 21205, USA.
Chem Biol. 2010 Jan 29;17(1):86-97. doi: 10.1016/j.chembiol.2009.12.014.
Methionine aminopeptidase (MetAP) is a metalloprotease that removes the N-terminal methionine during protein synthesis. To assess the importance of the two MetAPs in Mycobacterium tuberculosis, we overexpressed and purified each of the MetAPs to near homogeneity and showed that both were active as MetAP enzymes in vitro. We screened a library of 175,000 compounds against MtMetAP1c and identified 2,3-dichloro-1,4-naphthoquinone class of compounds as inhibitors of both MtMetAPs. It was found that the MtMetAP inhibitors were active against replicating and aged nongrowing M. tuberculosis. Overexpression of either MtMetAP1a or MtMetAP1c in M. tuberculosis conferred resistance of bacterial cells to the inhibitors. Moreover, knockdown of MtMetAP1a, but not MtMetAP1c, resulted in decreased viability of M. tuberculosis. These results suggest that MtMetAP1a is a promising target for developing antituberculosis agents.
甲硫氨酸氨肽酶（MetAP）是一种金属蛋白酶，在蛋白质合成过程中去除N端甲硫氨酸。为了评估两种MetAP在结核分枝杆菌中的重要性，我们将每种MetAP过表达并纯化至接近均一性，并表明两者在体外均作为MetAP酶具有活性。我们针对MtMetAP1c筛选了一个包含175,000种化合物的文库，并鉴定出2,3-二氯-1,4-萘醌类化合物是两种MtMetAP的抑制剂。发现MtMetAP抑制剂对正在复制和老化的非生长型结核分枝杆菌具有活性。在结核分枝杆菌中过表达MtMetAP1a或MtMetAP1c可使细菌细胞对抑制剂产生抗性。此外，敲低MtMetAP1a而非MtMetAP1c会导致结核分枝杆菌的活力下降。这些结果表明，MtMetAP1a是开发抗结核药物的一个有前景的靶点。
