Isabella Vincent M, Campbell Arthur J, Manchester John, Sylvester Mark, Nayar Asha S, Ferguson Keith E, Tommasi Ruben, Miller Alita A
Synlogic, 25 First Street, Cambridge, MA 02141, USA.
Stanley Center for Psychiatric Research, Broad Institute of Massachusetts Institute of Technology and Harvard University, 415 Main Street, Cambridge, MA 02142, USA.
Chem Biol. 2015 Apr 23;22(4):535-547. doi: 10.1016/j.chembiol.2015.03.018.
Understanding how compound penetration occurs across the complex cell walls of Gram-negative bacteria is one of the greatest challenges in discovering new drugs to treat the infections they cause. A combination of next-generation transposon sequencing, computational metadynamics simulations (CMDS), and medicinal chemistry was used to define genetic and structural elements involved in facilitated carbapenem entry into Pseudomonas aeruginosa. Here we show for the first time that these compounds are taken up not only by the major outer membrane channel OccD1 (also called OprD or PA0958) but also by a closely related channel OccD3 (OpdP or PA4501). Transport-mediating molecular interactions predicted by CMDS for these channels were first confirmed genetically, then used to guide the design of carbapenem analogs with altered uptake properties. These results bring us closer to the rational design of channel transmissibility and may ultimately lead to improved permeability of compounds across bacterial outer membranes.
了解化合物如何穿透革兰氏阴性菌复杂的细胞壁是发现治疗由它们引起的感染的新药面临的最大挑战之一。结合下一代转座子测序、计算元动力学模拟（CMDS）和药物化学来确定促进碳青霉烯类进入铜绿假单胞菌所涉及的遗传和结构元件。在这里，我们首次表明这些化合物不仅通过主要的外膜通道OccD1（也称为OprD或PA0958）进入，还通过密切相关的通道OccD3（OpdP或PA4501）进入。CMDS预测的这些通道的转运介导分子相互作用首先通过遗传学得到证实，然后用于指导具有改变摄取特性的碳青霉烯类似物的设计。这些结果使我们更接近通道传导性的合理设计，并最终可能提高化合物跨细菌外膜的通透性。
