Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase

被引:1
作者
Dirr, Larissa [1 ]
Cleeves, Sven [2 ,3 ]
Ramon Roth, Isabel [4 ]
Li, Linghui [1 ,10 ]
Fiebig, Timm [4 ]
Ve, Thomas [1 ]
Haeussler, Susanne [5 ,6 ,7 ,8 ]
Braun, Armin [2 ,3 ,9 ]
von Itzstein, Mark [1 ]
Fuehring, Jana I. [4 ]
机构
[1] Griffith Univ, Inst Glyc, Gold Coast Campus, Gold Coast, Qld, Australia
[2] Fraunhofer Int Consortium Antiinfect Res iCAIR, Fraunhofer Inst Toxicol & Expt Med ITEM, Hannover, Germany
[3] German Ctr Lung Res DZL, Biomed Res Endstage & Obstructive Lung Dis BREATH, Hannover, Germany
[4] Hannover Med Sch, Inst Clin Biochem, Hannover, Germany
[5] Helmholtz Ctr Infect Res, Dept Mol Bacteriol, Braunschweig, Germany
[6] TWINCORE, Inst Mol Bacteriol, Ctr Expt & Clin Infect Res, Hannover, Germany
[7] Copenhagen Univ Hosp, Dept Clin Microbiol, Rigshosp, Copenhagen, Denmark
[8] Hannover Med Sch, Cluster Excellence RESIST EXC 2155, Hannover, Germany
[9] Hannover Med Sch, Inst Immunol, Hannover, Germany
[10] Shenzhen Longhua High Sch Educ Grp, Shenzhen, Peoples R China
来源
MBIO | 2024年 / 15卷 / 04期
基金
英国医学研究理事会;
关键词
drug targets; Pseudomonas aeruginosa; protein structure-function; virulence factors; lipopolysaccharide; virulence; opportunistic infections; antibiotic resistance; lung infection; precision-cut lung slices; UDP-glucose pyrophosphorylase; galU; oligomerization; III PROTEIN SECRETION; LIPOPOLYSACCHARIDE CORE; OLIGOMERIZATION STATUS; MOLECULAR-CLONING; CRYSTAL-STRUCTURE; OUTER-MEMBRANE; GALU MUTANT; IDENTIFICATION; URIDYLYLTRANSFERASE; PNEUMONIA;
D O I
10.1128/mbio.02114-23
中图分类号
Q93 [微生物学];
学科分类号
071005 ; 100705 ;
摘要
Multidrug-resistant bacteria such as the opportunistic pathogen Pseudomonas aeruginosa, which causes life-threatening infections especially in immunocompromised individuals and cystic fibrosis patients, pose an increasing threat to public health. In the search for new treatment options, P. aeruginosa uridine diphosphate-glucose pyrophosphorylase (PaUGP) has been proposed as a novel drug target because it is required for the biosynthesis of important virulence factors and linked to pathogenicity in animal models. Here, we show that UGP-deficient P. aeruginosa exhibits severely reduced virulence against human lung tissue and cells, emphasizing the enzyme's suitability as a drug target. To establish a basis for the development of selective PaUGP inhibitors, we solved the product-bound crystal structure of tetrameric PaUGP and conducted a comprehensive structure-function analysis, identifying key residues at two different molecular interfaces that are essential for tetramer integrity and catalytic activity and demonstrating that tetramerization is pivotal for PaUGP function. Importantly, we show that part of the PaUGP oligomerization interface is uniquely conserved across bacterial UGPs but does not exist in the human enzyme, therefore representing an allosteric site that may be targeted to selectively inhibit bacterial UGPs. IMPORTANCE Infections with the opportunistic bacterial pathogen Pseudomonas aeruginosa are becoming increasingly difficult to treat due to multidrug resistance. Here, we show that the enzyme uridine diphosphate-glucose pyrophosphorylase (UGP) is involved in P. aeruginosa virulence toward human lung tissue and cells, making it a potential target for the development of new antibacterial drugs. Our exploration of P. aeruginosa (Pa)UGP structure-function relationships reveals that the activity of PaUGP depends on the formation of a tetrameric enzyme complex. We found that a molecular interface involved in tetramer formation is conserved in all bacterial UGPs but not in the human enzyme, and therefore hypothesize that it provides an ideal point of attack to selectively inhibit bacterial UGPs and exploit them as drug targets.
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页数:23
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