鉴定半胱氨酸代谢调节剂 (CymR) 衍生五肽作为金黄色葡萄球菌 O-乙酰基-l-丝氨酸巯基酶 (CysK) 的纳摩尔抑制剂。

IF 4 2区 医学 Q2 CHEMISTRY, MEDICINAL ACS Infectious Diseases Pub Date : 2025-01-10 Epub Date: 2024-12-20 DOI:10.1021/acsinfecdis.4c00832
Jordan L Pederick, Bethiney C Vandborg, Amir George, Hannah Bovermann, Jeffrey M Boyd, Joel S Freundlich, John B Bruning
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引用次数: 0

摘要

细菌半胱氨酸生物合成途径为抗生素佐剂的开发提供了动力。细菌半胱氨酸的生物合成通常由两种具有o -乙酰-l-丝氨酸巯基化酶(OASS)的酶CysK和CysM促进。在金黄色葡萄球菌中,存在一个单一的OASS同源物SaCysK。研究发现,敲除SaCysK可增加对氧化应激的敏感性,使其成为抑制剂开发的相关靶点。SaCysK还通过与丝氨酸乙酰转移酶(CysE)或半胱氨酸代谢转录调节因子(CymR)途径中的前一酶相互作用形成两种功能复合物。这些相互作用是通过将CysE或CymR的c端肽插入SaCysK的活性位点,抑制OASS活性而发生的,因此是开发SaCysK抑制剂的一个很好的起点。在这里,我们详细描述了CysE和cymr衍生的c端肽作为SaCysK抑制剂的特性。结合x射线晶体学、表面等离子体共振和酶抑制实验,确定cymr衍生的十肽与SaCysK形成广泛的相互作用,并作为一种有效的抑制剂(KD = 25 nM;IC50 = 180 nM),使其成为SaCysK抑制剂开发的有希望的先导物。为了了解这种高亲和力相互作用的决定因素,我们还研究了16种合理设计的肽的结构-活性关系。这表明CymR的c端五肽促进了与SaCysK的高亲和力相互作用,并且可以在不影响效力的情况下对五肽进行细微的结构修饰。最终,这项工作确定了CymR五肽作为开发针对SaCysK的抗生素佐剂的有前途的支架。
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Identification of Cysteine Metabolism Regulator (CymR)-Derived Pentapeptides as Nanomolar Inhibitors of Staphylococcus aureus O-Acetyl-l-serine Sulfhydrylase (CysK).

The pathway of bacterial cysteine biosynthesis is gaining traction for the development of antibiotic adjuvants. Bacterial cysteine biosynthesis is generally facilitated by two enzymes possessing O-acetyl-l-serine sulfhydrylases (OASS), CysK and CysM. In Staphylococcus aureus, there exists a single OASS homologue, SaCysK. Knockout of SaCysK was found to increase sensitivity to oxidative stress, making it a relevant target for inhibitor development. SaCysK also forms two functional complexes via interaction with the preceding enzyme in the pathway serine acetyltransferase (CysE) or the transcriptional regulator of cysteine metabolism (CymR). These interactions occur through insertion of a C-terminal peptide of CysE or CymR into the active site of SaCysK, inhibiting OASS activity, and therefore represent an excellent starting point for developing SaCysK inhibitors. Here, we detail the characterization of CysE and CymR-derived C-terminal peptides as inhibitors of SaCysK. Using a combination of X-ray crystallography, surface plasmon resonance, and enzyme inhibition assays, it was determined that the CymR-derived decapeptide forms extensive interactions with SaCysK and acts as a potent inhibitor (KD = 25 nM; IC50 = 180 nM), making it a promising lead for the development of SaCysK inhibitors. To understand the determinants of this high-affinity interaction, the structure-activity relationships of 16 rationally designed peptides were also investigated. This identified that the C-terminal pentapeptide of CymR facilitates the high-affinity interaction with SaCysK and that subtle structural modification of the pentapeptide is possible without impacting potency. Ultimately, this work identified CymR pentapeptides as a promising scaffold for the development of antibiotic adjuvants targeting SaCysK.

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来源期刊
ACS Infectious Diseases
ACS Infectious Diseases CHEMISTRY, MEDICINALINFECTIOUS DISEASES&nb-INFECTIOUS DISEASES
CiteScore
9.70
自引率
3.80%
发文量
213
期刊介绍: ACS Infectious Diseases will be the first journal to highlight chemistry and its role in this multidisciplinary and collaborative research area. The journal will cover a diverse array of topics including, but not limited to: * Discovery and development of new antimicrobial agents — identified through target- or phenotypic-based approaches as well as compounds that induce synergy with antimicrobials. * Characterization and validation of drug target or pathways — use of single target and genome-wide knockdown and knockouts, biochemical studies, structural biology, new technologies to facilitate characterization and prioritization of potential drug targets. * Mechanism of drug resistance — fundamental research that advances our understanding of resistance; strategies to prevent resistance. * Mechanisms of action — use of genetic, metabolomic, and activity- and affinity-based protein profiling to elucidate the mechanism of action of clinical and experimental antimicrobial agents. * Host-pathogen interactions — tools for studying host-pathogen interactions, cellular biochemistry of hosts and pathogens, and molecular interactions of pathogens with host microbiota. * Small molecule vaccine adjuvants for infectious disease. * Viral and bacterial biochemistry and molecular biology.
期刊最新文献
Decoding the Role of Antimicrobial Peptides in the Fight against Mycobacterium tuberculosis. Metabolic Flexibility and Essentiality of the Tricarboxylic Acid Cycle in Plasmodium. Elucidation of the Glycan Structure of the b-type Flagellin of Pseudomonas aeruginosa PAO1. Advantages and Challenges of Using Antimicrobial Peptides in Synergism with Antibiotics for Treating Multidrug-Resistant Bacteria. Building Spatiotemporal Understanding of Mycobacterium tuberculosis-Host Interactions.
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