pirin 蛋白和 2-ketoglutarate:脆弱拟杆菌新陈代谢中铁红蛋白氧化还原酶同源物的新功能及其对甲硝唑和阿米西利抗菌药敏感性的影响。

IF 3.9 3区 生物学 Q2 MICROBIOLOGY
MicrobiologyOpen Pub Date : 2024-08-07 DOI:10.1002/mbo3.1429
Andrea M. Gough, Anita C. Parker, Patricia J. O'Bryan, Terence R. Whitehead, Sourav Roy, Brandon L. Garcia, Paul S. Hoffman, C. Jeffrey Smith, Edson R. Rocha
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引用次数: 0

摘要

人们对厌氧病原体脆弱拟杆菌(Bacteroides fragilis)的中心代谢和发酵途径如何调控其对抗菌素的敏感性的认识仍不完整。我们的研究发现,脆弱拟杆菌编码两个铁依赖性、氧化还原敏感性调控 pirin 蛋白基因 pir1 和 pir2,当暴露于氧气和在铁限制条件下生长时,这些基因的 mRNA 表达增加。这些蛋白(Pir1 和 Pir2)会影响短链脂肪酸的产生,并改变厌氧菌对甲硝唑和丙酮酸铁氧还蛋白氧化还原酶新抑制剂 amixicile 的敏感性。我们通过双杂交系统检测证实,Pir1 和 Pir2 与这种氧化还原酶直接相互作用。此外,利用 AlphaFold2 进行的结构分析预测,Pir1 和 Pir2 与几种中央代谢酶有稳定的相互作用,包括 2-酮戊二酸:铁氧还蛋白氧化还原酶 Kor1AB 和 Kor2CDAEBG。我们利用一系列代谢突变体和电子传递链抑制剂证明了细菌代谢对甲硝唑和阿米西利敏感性的广泛影响。我们还表明,在腹腔内感染的实验模型中,氨苯砜是一种有效的抗菌剂。我们的研究发现,kor2AEBG 基因是生长所必需的,具有双重功能,包括通过反向 TCA 循环形成 2-酮戊二酸。然而,在添加磷脂或脂肪酸后,绕过 Kor2AEBG 功能的代谢活动仍未确定。总之,我们的研究为了解脆弱拟杆菌的中心代谢及其受 pirin 蛋白的调控提供了新的视角,这些视角可用于开发新的窄谱抗菌药物。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

New functions of pirin proteins and a 2-ketoglutarate: Ferredoxin oxidoreductase ortholog in Bacteroides fragilis metabolism and their impact on antimicrobial susceptibility to metronidazole and amixicile

New functions of pirin proteins and a 2-ketoglutarate: Ferredoxin oxidoreductase ortholog in Bacteroides fragilis metabolism and their impact on antimicrobial susceptibility to metronidazole and amixicile

The understanding of how central metabolism and fermentation pathways regulate antimicrobial susceptibility in the anaerobic pathogen Bacteroides fragilis is still incomplete. Our study reveals that B. fragilis encodes two iron-dependent, redox-sensitive regulatory pirin protein genes, pir1 and pir2. The mRNA expression of these genes increases when exposed to oxygen and during growth in iron-limiting conditions. These proteins, Pir1 and Pir2, influence the production of short-chain fatty acids and modify the susceptibility to metronidazole and amixicile, a new inhibitor of pyruvate: ferredoxin oxidoreductase in anaerobes. We have demonstrated that Pir1 and Pir2 interact directly with this oxidoreductase, as confirmed by two-hybrid system assays. Furthermore, structural analysis using AlphaFold2 predicts that Pir1 and Pir2 interact stably with several central metabolism enzymes, including the 2-ketoglutarate:ferredoxin oxidoreductases Kor1AB and Kor2CDAEBG. We used a series of metabolic mutants and electron transport chain inhibitors to demonstrate the extensive impact of bacterial metabolism on metronidazole and amixicile susceptibility. We also show that amixicile is an effective antimicrobial against B. fragilis in an experimental model of intra-abdominal infection. Our investigation led to the discovery that the kor2AEBG genes are essential for growth and have dual functions, including the formation of 2-ketoglutarate via the reverse TCA cycle. However, the metabolic activity that bypasses the function of Kor2AEBG following the addition of phospholipids or fatty acids remains undefined. Overall, our study provides new insights into the central metabolism of B. fragilis and its regulation by pirin proteins, which could be exploited for the development of new narrow-spectrum antimicrobials in the future.

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来源期刊
MicrobiologyOpen
MicrobiologyOpen MICROBIOLOGY-
CiteScore
8.00
自引率
0.00%
发文量
78
审稿时长
20 weeks
期刊介绍: MicrobiologyOpen is a peer reviewed, fully open access, broad-scope, and interdisciplinary journal delivering rapid decisions and fast publication of microbial science, a field which is undergoing a profound and exciting evolution in this post-genomic era. The journal aims to serve the research community by providing a vehicle for authors wishing to publish quality research in both fundamental and applied microbiology. Our goal is to publish articles that stimulate discussion and debate, as well as add to our knowledge base and further the understanding of microbial interactions and microbial processes. MicrobiologyOpen gives prompt and equal consideration to articles reporting theoretical, experimental, applied, and descriptive work in all aspects of bacteriology, virology, mycology and protistology, including, but not limited to: - agriculture - antimicrobial resistance - astrobiology - biochemistry - biotechnology - cell and molecular biology - clinical microbiology - computational, systems, and synthetic microbiology - environmental science - evolutionary biology, ecology, and systematics - food science and technology - genetics and genomics - geobiology and earth science - host-microbe interactions - infectious diseases - natural products discovery - pharmaceutical and medicinal chemistry - physiology - plant pathology - veterinary microbiology We will consider submissions across unicellular and cell-cluster organisms: prokaryotes (bacteria, archaea) and eukaryotes (fungi, protists, microalgae, lichens), as well as viruses and prions infecting or interacting with microorganisms, plants and animals, including genetic, biochemical, biophysical, bioinformatic and structural analyses. The journal features Original Articles (including full Research articles, Method articles, and Short Communications), Commentaries, Reviews, and Editorials. Original papers must report well-conducted research with conclusions supported by the data presented in the article. We also support confirmatory research and aim to work with authors to meet reviewer expectations. MicrobiologyOpen publishes articles submitted directly to the journal and those referred from other Wiley journals.
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