Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase
ABSTRACTMultidrug-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. a...
| Main Authors: | , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2024-04-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mbio.02114-23 |
| _version_ | 1797215319795695616 |
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| author | Larissa Dirr Sven Cleeves Isabel Ramón Roth Linghui Li Timm Fiebig Thomas Ve Susanne Häussler Armin Braun Mark von Itzstein Jana I. Führing |
| author_facet | Larissa Dirr Sven Cleeves Isabel Ramón Roth Linghui Li Timm Fiebig Thomas Ve Susanne Häussler Armin Braun Mark von Itzstein Jana I. Führing |
| author_sort | Larissa Dirr |
| collection | DOAJ |
| description | ABSTRACTMultidrug-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.IMPORTANCEInfections 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. |
| first_indexed | 2024-04-24T11:28:11Z |
| format | Article |
| id | doaj.art-d8d00c4b95764f78a4b8670e9a0d7496 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-04-24T11:28:11Z |
| publishDate | 2024-04-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-d8d00c4b95764f78a4b8670e9a0d74962024-04-10T13:01:15ZengAmerican Society for MicrobiologymBio2150-75112024-04-0115410.1128/mbio.02114-23Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylaseLarissa Dirr0Sven Cleeves1Isabel Ramón Roth2Linghui Li3Timm Fiebig4Thomas Ve5Susanne Häussler6Armin Braun7Mark von Itzstein8Jana I. Führing9Institute for Glycomics, Gold Coast Campus, Griffith University, Gold Coast, Queensland, AustraliaFraunhofer Institute for Toxicology and Experimental Medicine ITEM, Member of Fraunhofer International Consortium for Anti-Infective Research (iCAIR), Hannover, GermanyInstitute of Clinical Biochemistry, Hannover Medical School, Hannover, GermanyInstitute for Glycomics, Gold Coast Campus, Griffith University, Gold Coast, Queensland, AustraliaInstitute of Clinical Biochemistry, Hannover Medical School, Hannover, GermanyInstitute for Glycomics, Gold Coast Campus, Griffith University, Gold Coast, Queensland, AustraliaDepartment of Molecular Bacteriology, Helmholtz Centre for Infection Research, Braunschweig, GermanyFraunhofer Institute for Toxicology and Experimental Medicine ITEM, Member of Fraunhofer International Consortium for Anti-Infective Research (iCAIR), Hannover, GermanyInstitute for Glycomics, Gold Coast Campus, Griffith University, Gold Coast, Queensland, AustraliaInstitute of Clinical Biochemistry, Hannover Medical School, Hannover, GermanyABSTRACTMultidrug-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.IMPORTANCEInfections 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.https://journals.asm.org/doi/10.1128/mbio.02114-23drug targetsPseudomonas aeruginosaprotein structure-functionvirulence factorslipopolysaccharidevirulence |
| spellingShingle | Larissa Dirr Sven Cleeves Isabel Ramón Roth Linghui Li Timm Fiebig Thomas Ve Susanne Häussler Armin Braun Mark von Itzstein Jana I. Führing Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase mBio drug targets Pseudomonas aeruginosa protein structure-function virulence factors lipopolysaccharide virulence |
| title | Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase |
| title_full | Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase |
| title_fullStr | Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase |
| title_full_unstemmed | Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase |
| title_short | Tetramerization is essential for the enzymatic function of the Pseudomonas aeruginosa virulence factor UDP-glucose pyrophosphorylase |
| title_sort | tetramerization is essential for the enzymatic function of the pseudomonas aeruginosa virulence factor udp glucose pyrophosphorylase |
| topic | drug targets Pseudomonas aeruginosa protein structure-function virulence factors lipopolysaccharide virulence |
| url | https://journals.asm.org/doi/10.1128/mbio.02114-23 |
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