The long chain fatty acid-CoA ligase VraA plays a regulatory role in vancomycin resistance in Staphylococcus aureus

IF 1.8 4区生物学 Q4 BIOCHEMICAL RESEARCH METHODS

Protein expression and purification Pub Date : 2026-02-01 Epub Date: 2025-10-21 DOI:10.1016/j.pep.2025.106838

Dafeng Liu , Na Li

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引用次数: 0

Abstract

The pathogen Staphylococcus aureus threatens clinical and public health. Vancomycin, a former last-resort treatment, has waning efficacy against resistant strains (VISA) that alter their cell envelope using enzymes like VraA. However, the specific functional mechanism of VraA in VISA is still not understood. Here, we successfully obtained monomeric VraA and found that oleate is the optimal substrate for VraA. VraA was characterized as a peripheral membrane protein with a hydrodynamic radius of 5.7 ± 0.3 nm. Disruption of VraA significantly increased S. aureus susceptibility to vancomycin, as demonstrated by minimum inhibitory concentration (MIC) assays and RT-qPCR. The structural model of VraA predicted using AlphaFold2 was refined via ModRefiner, and active site residues were identified. Our findings underscore the pivotal role of VraA in mediating vancomycin resistance in S. aureus, providing valuable insights for developing targeted therapies against this pathogen.

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长链脂肪酸-辅酶a连接酶VraA在金黄色葡萄球菌万古霉素耐药过程中起调节作用。

病原菌金黄色葡萄球菌威胁着临床和公众健康。万古霉素曾是一种最后的治疗手段，但它对利用VraA等酶改变细胞包膜的耐药菌株（VISA）的疗效正在减弱。然而，VraA在VISA中的具体作用机制尚不清楚。在这里，我们成功地获得了单体VraA，并发现油酸酯是VraA的最佳底物。VraA为外周膜蛋白，水动力半径为5.7±0.3 nm。最低抑制浓度（MIC）测定和RT-qPCR结果表明，破坏VraA可显著增加金黄色葡萄球菌对万古霉素的敏感性。通过ModRefiner对AlphaFold2预测的VraA结构模型进行细化，并鉴定出活性位点残基。我们的发现强调了VraA在介导金黄色葡萄球菌万古霉素耐药性中的关键作用，为开发针对该病原体的靶向治疗提供了有价值的见解。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Protein expression and purification 生物-生化研究方法

CiteScore

3.70

自引率

6.20%

发文量

120

审稿时长

32 days

期刊介绍： Protein Expression and Purification is an international journal providing a forum for the dissemination of new information on protein expression, extraction, purification, characterization, and/or applications using conventional biochemical and/or modern molecular biological approaches and methods, which are of broad interest to the field. The journal does not typically publish repetitive examples of protein expression and purification involving standard, well-established, methods. However, exceptions might include studies on important and/or difficult to express and/or purify proteins and/or studies that include extensive protein characterization, which provide new, previously unpublished information.