Fluorescent Benzothiazinone Analogues Efficiently and Selectively Label Dpre1 in Mycobacteria and Actinobacteria,ACS Chemical Biology

当前位置： X-MOL 学术 › ACS Chem. Biol. › 论文详情

Our official English website, www.x-mol.net, welcomes your feedback! (Note: you will need to create a separate account there.)

Fluorescent Benzothiazinone Analogues Efficiently and Selectively Label Dpre1 in Mycobacteria and Actinobacteria
ACS Chemical Biology ( IF 3.5 ) Pub Date : 2018-10-05 00:00:00 , DOI: 10.1021/acschembio.8b00790
Raphael Sommer ₁ , João Neres _{1,

2} , Jérémie Piton _{1,

2} , Neeraj Dhar _{1,

2} , Astrid van der Sar ₃ , Raju Mukherjee _{1,

2} , Thierry Laroche ₄ , Paul J. Dyson ₅ , John D. McKinney _{1,

2} , Wilbert Bitter ₃ , Vadim Makarov ₆ , Stewart T. Cole _{1,

2}

Affiliation

Benzothiazinones (BTZ) are highly potent bactericidal inhibitors of mycobacteria and the lead compound, BTZ043, and the optimized drug candidate, PBTZ169, have potential for the treatment of tuberculosis. Here, we exploited the tractability of the BTZ scaffold by attaching a range of fluorophores to the 2-substituent of the BTZ ring via short linkers. We show by means of fluorescence imaging that the most advanced derivative, JN108, is capable of efficiently labeling its target, the essential flavoenzyme DprE1, both in cell-free extracts and after purification as well as in growing cells of different actinobacterial species. DprE1 displays a polar localization in Mycobacterium tuberculosis, M. marinum, M. smegmatis, and Nocardia farcinica but not in Corynebacterium glutamicum. Finally, mutation of the cysteine residue in DprE1 in these species, to which BTZ covalently binds, abolishes completely the interaction with JN108, thereby highlighting the specificity of this fluorescent probe.

中文翻译：

荧光苯并噻嗪酮类似物有效，选择性地标记分枝杆菌和放线菌中的Dpre1。

苯并噻嗪酮（BTZ）是分枝杆菌的高效杀菌抑制剂，其主要化合物BTZ043和优化的候选药物PBTZ169在治疗结核病方面具有潜力。在这里，我们通过短连接子将一系列荧光团连接到BTZ环的2位取代基上，从而开发了BTZ支架的可延展性。我们通过荧光成像显示，最先进的衍生物JN108能够在无细胞提取物中，纯化后以及在不同放线菌物种的生长细胞中有效地标记其靶标，必需的黄素酶DprE1。DprE1显示极性定位结核分枝杆菌，海分枝杆菌，耻垢分枝杆菌，和诺卡farcinica但不能在谷氨酸棒状杆菌。最后，BTZ共价结合的这些物种中DprE1中半胱氨酸残基的突变完全消除了与JN108的相互作用，从而突出了这种荧光探针的特异性。

更新日期：2018-10-05

点击分享查看原文

点击收藏

阅读更多本刊最新论文本刊介绍/投稿指南11