Unraveling the Protein Targets of Vancomycin in Living S. aureus and E. faecalis Cells
Unraveling the Protein Targets of Vancomycin in Living S. aureus and E. faecalis Cells
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DOI：
10.1021/ja2039979
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发表时间：
2011-08-10
影响因子：
15
通讯作者：
Sieber, Stephan A.
中科院分区：
文献类型：
--
作者：
Eirich, Juergen;Orth, Ronald;Sieber, Stephan A.
万古霉素是一种有效的糖肽抗生素，已进化为与新生肽聚糖的D-丙氨酸-D-丙氨酸二肽末端特异性结合。虽然这种作用模式已经确立，但一些研究表明，万古霉素及其类似物利用了非规范的靶点。为了解决临床相关的金黄色葡萄球菌和粪肠球菌的所有万古霉素靶点，我们开发了一系列以万古霉素为基础的小分子光亲和探针。蛋白质组学分析揭示了两个以前未知的万古霉素靶点的特定标记，这两个靶点可能有助于其抗生素活性。对主要葡萄球菌自溶素AT1的特异性抑制证实了先前的观察结果，即万古霉素通过与自溶机制相互作用改变金黄色葡萄球菌的细胞形态。此外，在粪肠球菌中，万古霉素光探针与ABC转运蛋白特异性结合，这可能会阻碍糖和肽等必要营养物质的吸收。对活细胞中这两个突出的膜靶标的标记揭示了一种迄今未被探索的万古霉素结合和抑制模式，该模式可能允许合理设计具有更高活性的变体。
Vancomycin is a potent glycopeptide antibiotic that has evolved to specifically bind to the D-Ala-D-Ala dipeptide termini of nascent peptidoglycans. Although this mode of action is well established, several studies indicate that vancomycin and analogues exploit noncanonical target sites. In order to address all vancomycin targets in clinically relevant Staphylococcus aureus and Enterococcus faecalis strains we developed a series of small-molecule photoaffinity probes based on vancomycin. Proteomic profiling revealed the specific labeling of two previously unknown vancomycin targets that are likely to contribute to its antibiotic activity. The specific inhibition of the major staphylococcal autolysin Atl confirms previous observations that vancomycin alters S. aureus cell morphology by interaction with the autolytic machinery. Moreover, in E. faecalis the vancomycin photoprobe specifically binds to an ABC transporter protein, which likely impedes the uptake of essential nutrients such as sugars and peptides. The labeling of these two prominent membrane targets in living cells reveals a thus far unexplored mode of vancomycin binding and inhibition that could allow a rational design of variants with improved activity.