Identification and characterization of the functional tetrameric UDP-glucose pyrophosphorylase from Klebsiella pneumoniae.

IF 5.1 1区 生物学 Q1 MICROBIOLOGY
mBio Pub Date : 2024-12-20 DOI:10.1128/mbio.02071-24
Isabel Ramón Roth, Pavel Kats, Timm Fiebig, Françoise Routier, Roman Fedorov, Larissa Dirr, Jana I Führing
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引用次数: 0

Abstract

In all kingdoms of life, the enzyme uridine diphosphate-glucose pyrophosphorylase (UGP) occupies a central role in metabolism, as its reaction product uridine diphosphate-glucose (UDP-Glc) is involved in various crucial cellular processes. Pathogens, including fungi, parasites, and bacteria, depend on UGP for the synthesis of virulence factors; in particular, various bacterial species utilize UDP-Glc and its derivatives for the synthesis of lipopolysaccharides, capsular polysaccharides, and biofilm exopolysaccharides. UGPs have, therefore, gained attention as anti-bacterial drug target candidates, prompting us to study their structure-function relationships to provide a basis for the rational development of specific inhibitors. UGP function is tied to its oligomeric state, and the majority of bacterial homologs have been described as tetramers encoded by the galU gene. Uniquely, enterobacterial species harbor a second gene, galF, encoding a protein with high homology to UGP, whose function is somewhat controversial. Here, we show that the galF gene of the opportunistic pathogen Klebsiella pneumoniae encodes a dimeric protein that has lost UGP activity, likely due to a combination of active site mutations and an inability to tetramerize, whereas the functional K. pneumoniae UGP, encoded by galU, is an active tetramer. Our AlphaFold-assisted structure-function relationship studies underline that tetramerization is essential for bacterial UGP function and is facilitated by a common mechanism utilizing conserved key residues. Targeting the respective molecular interfaces, which are absent in human UGP, could provide a means of selectively inhibiting the bacterial virulence factor UGP and potentially rendering pathogenic species avirulent.IMPORTANCEThe enzyme uridine diphosphate-glucose pyrophosphorylase (UGP) is important for the virulence of bacterial pathogens and, therefore, a potential drug target. In this study, we identify the gene encoding the functional UGP in Klebsiella pneumoniae, a bacterium notoriously causing severe antibiotic-resistant infections in humans, and reveal structural and functional features that may aid in the development of new antibiotics.

在所有生物界中,尿苷二磷酸-葡萄糖焦磷酸化酶(UGP)在新陈代谢中都扮演着核心角色,因为它的反应产物尿苷二磷酸-葡萄糖(UDP-Glc)参与了各种关键的细胞过程。包括真菌、寄生虫和细菌在内的病原体依赖 UGP 合成毒力因子,特别是各种细菌利用 UDP-Glc 及其衍生物合成脂多糖、胶囊多糖和生物膜外多糖。因此,UGPs 作为抗菌药物的候选靶标而备受关注,促使我们研究其结构与功能的关系,为合理开发特异性抑制剂奠定基础。UGP 的功能与其寡聚状态有关,大多数细菌同源物都是由 galU 基因编码的四聚体。与众不同的是,肠杆菌中还存在第二个基因 galF,它编码一种与 UGP 高度同源的蛋白质,但其功能尚存在争议。在这里,我们发现机会性病原体肺炎克雷伯菌的 galF 基因编码的是一种失去 UGP 活性的二聚体蛋白,这可能是由于活性位点突变和无法四聚化的共同作用造成的,而由 galU 编码的功能性肺炎克雷伯菌 UGP 则是一种活性四聚体。我们的 AlphaFold 辅助结构-功能关系研究强调,四聚体化对细菌 UGP 的功能至关重要,并通过利用保守的关键残基的共同机制来促进四聚体化。针对人类 UGP 中不存在的相应分子界面,可以提供一种选择性抑制细菌毒力因子 UGP 的方法,并有可能使致病物种变得无毒。重要意义尿苷二磷酸-葡萄糖焦磷酸酶(UGP)对细菌病原体的毒力非常重要,因此是一种潜在的药物靶标。在这项研究中,我们确定了肺炎克雷伯氏菌(一种导致人类严重抗生素耐药性感染的臭名昭著的细菌)中编码功能性 UGP 的基因,并揭示了其结构和功能特征,这些特征可能有助于新抗生素的开发。
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来源期刊
mBio
mBio MICROBIOLOGY-
CiteScore
10.50
自引率
3.10%
发文量
762
审稿时长
1 months
期刊介绍: mBio® is ASM''s first broad-scope, online-only, open access journal. mBio offers streamlined review and publication of the best research in microbiology and allied fields.
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