脂多糖转运抑制剂的发现：活性测定方法的建立
Development of an activity assay for discovery of inhibitors of lipopolysaccharide transport.
机构信息
Department of Chemistry and Chemical Biology, Harvard University, Cambridge, Massachusetts 01238, USA.
出版信息
J Am Chem Soc. 2010 Mar 3;132(8):2518-9. doi: 10.1021/ja910361r.
Abstract
The outer membrane of gram-negative bacteria contains an outer leaflet composed of lipopolysaccharide (LPS) that is transported to this location by a pathway that is essential for viability. It has been suggested that inhibitors of this pathway could be useful antibiotics. Herein we reconstitute the activity of the ATPase component (LptB) of the ABC transporter that initiates LPS transport and assembly. We developed a high-throughput assay and screened a library of kinase inhibitors against LptB. We identified two classes of ATP-competitive inhibitors. These are the first inhibitors of the ATPase component of any bacterial ABC transporter. The small-molecule inhibitors will be very useful tools for further biochemical studies of the proteins involved in LPS transport and assembly.
摘要
革兰氏阴性细菌的外膜含有一个由脂多糖（LPS）组成的外叶，该 LPS 通过对生存至关重要的途径运输到该位置。有人提出，该途径的抑制剂可能是有用的抗生素。在此，我们重建了启动 LPS 运输和组装的 ABC 转运体的 ATP 酶组件（LptB）的活性。我们开发了一种高通量测定法，并针对 LptB 筛选了激酶抑制剂文库。我们确定了两类 ATP 竞争性抑制剂。这些是任何细菌 ABC 转运体的 ATP 酶组件的第一批抑制剂。这些小分子抑制剂将是进一步研究 LPS 运输和组装所涉及的蛋白质的生化研究的非常有用的工具。
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