The rapid emergence of drug resistant Staphylococcus aureus (S. aureus) poses a serious threat to public health globally. Silver (Ag)-based antimicrobials are promising to combat antibiotic resistant S. aureus, yet their molecular targets are largely elusive. Herein, we separate and identify 38 authentic Ag⁺-binding proteins in S. aureus at the whole-cell scale. We then capture the molecular snapshot on the dynamic action of Ag⁺ against S. aureus and further validate that Ag⁺ could inhibit a key target 6-phosphogluconate dehydrogenase through binding to catalytic His185 by X-ray crystallography. Significantly, the multi-target mode of action of Ag⁺ (and nanosilver) endows its sustainable antimicrobial efficacy, leading to enhanced efficacy of conventional antibiotics and resensitization of MRSA to antibiotics. Our study resolves the long-standing question of the molecular targets of silver in S. aureus and offers insights into the sustainable bacterial susceptibility of silver, providing a potential approach for combating antimicrobial resistance.

耐药金黄色葡萄球菌( S. aureus ) 的迅速出现对全球公共卫生构成严重威胁。基于银 (Ag) 的抗菌剂有望对抗抗生素耐药性金黄色葡萄球菌，但它们的分子靶点在很大程度上难以捉摸。在此，我们在全细胞范围内分离并鉴定了金黄色葡萄球菌中的38 种真正的 Ag ⁺结合蛋白。^{然后，我们捕捉到 Ag +}对金黄色葡萄球菌的动态作用的分子快照，并进一步验证 Ag ⁺可以通过 X 射线晶体学与催化 His185 结合来抑制关键靶标 6-磷酸葡萄糖酸脱氢酶。^{重要的是，Ag +} （和纳米银）的多靶点作用模式赋予其可持续的抗菌功效，从而提高常规抗生素的功效和 MRSA 对抗生素的重新敏感。我们的研究解决了金黄色葡萄球菌中银的分子靶标的长期问题，并提供了对银的可持续细菌敏感性的见解，为对抗抗菌素耐药性提供了一种潜在的方法。