A Target-Based Whole Cell Screen Approach To Identify Potential Inhibitors of Mycobacterium tuberculosis Signal Peptidase.
A Target-Based Whole Cell Screen Approach To Identify Potential Inhibitors of Mycobacterium tuberculosis Signal Peptidase.
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DOI：
10.1021/acsinfecdis.6b00075
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发表时间：
2016-12-09
影响因子：
5.3
通讯作者：
Parish, Tanya
中科院分区：
文献类型：
--
作者：
Bonnett, Shilah A.;Ollinger, Juliane;Chandrasekera, Susantha;Florio, Stephanie;O'Malley, Theresa;Files, Megan;Jee, Jo-Ann;Ahn, James;Casey, Allen;Ovechkina, Yulia;Roberts, David;Korkegian, Aaron;Parish, Tanya
The general secretion (Sec) pathway is a conserved essential pathway in bacteria and is the primary route of protein export across the cytoplasmic membrane. During protein export, the signal peptidase LepB catalyzes the cleavage of the signal peptide and subsequent release of mature proteins into the extracellular space. We developed a target-based whole cell assay to screen for potential inhibitors of LepB, the sole signal peptidase in Mycobacterium tuberculosis, using a strain engineered to underexpress LepB (LepB-UE). We screened 72,000 compounds against both the Lep-UE and wild-type (wt) strains. We identified the phenylhydrazone (PHY) series as having higher activity against the LepB-UE strain. We conducted a limited structure–activity relationship determination around a representative PHY compound with differential activity (MICs of 3.0 μM against the LepB-UE strain and 18 μM against the wt); several analogues were less potent against the LepB overexpressing strain. A number of chemical modifications around the hydrazone moiety resulted in improved potency. Inhibition of LepB activity was observed for a number of compounds in a biochemical assay using cell membrane fraction derived from M. tuberculosis. Compounds did not increase cell permeability, dissipate membrane potential, or inhibit an unrelated mycobacterial enzyme, suggesting a specific mode of action related to the LepB secretory mechanism.
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影响因子：
3.7
作者：
Kumar A;Zhang M;Zhu L;Liao RP;Mutai C;Hafsat S;Sherman DR;Wang MW
通讯作者：
Wang MW
影响因子：
3.1
作者：
Feltcher ME;Sullivan JT;Braunstein M
通讯作者：
Braunstein M
影响因子：
64.5
作者：
KOSHLAND, D;BOTSTEIN, D
通讯作者：
BOTSTEIN, D
影响因子：
3.5
作者：
Imramovsky, Ales;Polanc, Slovenko;Kaustova, Jarmila
通讯作者：
Kaustova, Jarmila
影响因子：
3.2
作者：
BLACK, MT
通讯作者：
BLACK, MT