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Targeting Mannitol metabolism as an alternative antimicrobial strategy based on the structure-function study of Mannitol-1-phosphate dehydrogenase in Staphylococcus aureus

Targeting Mannitol metabolism as an alternative antimicrobial strategy based on the structure-function study of Mannitol-1-phosphate dehydrogenase in Staphylococcus aureus
Targeting Mannitol metabolism as an alternative antimicrobial strategy based on the structure-function study of Mannitol-1-phosphate dehydrogenase in Staphylococcus aureus
Mannitol-1-phosphate dehydrogenase (M1PDH) is a key enzyme in Staphylococcus aureus mannitol metabolism, but its roles in pathophysiological settings have not been established. We performed comprehensive structure-function analysis of M1PDH from S. aureus USA300, a strain of community-associated methicillin-resistant S. aureus, to evaluate its roles in cell viability and virulence under pathophysiological conditions. On the basis of our results, we propose M1PDH as a potential antibacterial target. In vitro cell viability assessment of ΔmtlD knockout and complemented strains confirmed that M1PDH is essential to endure pH, high-salt, and oxidative stress and thus that M1PDH is required for preventing osmotic burst by regulating pressure potential imposed by mannitol. The mouse infection model also verified that M1PDH is essential for bacterial survival during infection. To further support the use of M1PDH as an antibacterial target, we identified dihydrocelastrol (DHCL) as a competitive inhibitor of S. aureus M1PDH (SaM1PDH) and confirmed that DHCL effectively reduces bacterial cell viability during host infection. To explain physiological functions of SaM1PDH at the atomic level, the crystal structure of SaM1PDH was determined at 1.7-Å resolution. Structure-based mutation analyses and DHCL molecular docking to the SaM1PDH active site followed by functional assay identified key residues in the active site and provided the action mechanism of DHCL. Collectively, we propose SaM1PDH as a target for antibiotic development based on its physiological roles with the goals of expanding the repertory of antibiotic targets to fight antimicrobial resistance and providing essential knowledge for developing potent inhibitors of SaM1PDH based on structure-function studies.IMPORTANCE Due to the shortage of effective antibiotics against drug-resistant Staphylococcus aureus, new targets are urgently required to develop next-generation antibiotics. We investigated mannitol-1-phosphate dehydrogenase of S. aureus USA300 (SaM1PDH), a key enzyme regulating intracellular mannitol levels, and explored the possibility of using SaM1PDH as a target for developing antibiotic. Since mannitol is necessary for maintaining the cellular redox and osmotic potential, the homeostatic imbalance caused by treatment with a SaM1PDH inhibitor or knockout of the gene encoding SaM1PDH results in bacterial cell death through oxidative and/or mannitol-dependent cytolysis. We elucidated the molecular mechanism of SaM1PDH and the structural basis of substrate and inhibitor recognition by enzymatic and structural analyses of SaM1PDH. Our results strongly support the concept that targeting of SaM1PDH represents an alternative strategy for developing a new class of antibiotics that cause bacterial cell death not by blocking key cellular machinery but by inducing cytolysis and reducing stress tolerance through inhibition of the mannitol pathway.
2150-7511
e02660-18
Nguyen, Thanh
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Kim, Truc
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Ta, Hai Minh
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Yeo, Won Sik
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Choi, Jongkeun
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Mizar, Pushpak
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Lee, Seung Seo
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Bae, Taeok
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Chaurasia, Akhilesh Kumar
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Kim, Kyeong Kyu
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Nguyen, Thanh
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Kim, Truc
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Ta, Hai Minh
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Yeo, Won Sik
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Choi, Jongkeun
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Mizar, Pushpak
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Lee, Seung Seo
ee34fa26-5fb6-48c8-80c2-1f13ec4ccceb
Bae, Taeok
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Chaurasia, Akhilesh Kumar
8f90122d-8b19-4545-9113-03ddac316b66
Kim, Kyeong Kyu
a7757cb7-065e-4e76-a669-27d2108bef97

Nguyen, Thanh, Kim, Truc, Ta, Hai Minh, Yeo, Won Sik, Choi, Jongkeun, Mizar, Pushpak, Lee, Seung Seo, Bae, Taeok, Chaurasia, Akhilesh Kumar and Kim, Kyeong Kyu (2019) Targeting Mannitol metabolism as an alternative antimicrobial strategy based on the structure-function study of Mannitol-1-phosphate dehydrogenase in Staphylococcus aureus. mBio, 10 (4), e02660-18. (doi:10.1128/mBio.02660-18).

Record type: Article

Abstract

Mannitol-1-phosphate dehydrogenase (M1PDH) is a key enzyme in Staphylococcus aureus mannitol metabolism, but its roles in pathophysiological settings have not been established. We performed comprehensive structure-function analysis of M1PDH from S. aureus USA300, a strain of community-associated methicillin-resistant S. aureus, to evaluate its roles in cell viability and virulence under pathophysiological conditions. On the basis of our results, we propose M1PDH as a potential antibacterial target. In vitro cell viability assessment of ΔmtlD knockout and complemented strains confirmed that M1PDH is essential to endure pH, high-salt, and oxidative stress and thus that M1PDH is required for preventing osmotic burst by regulating pressure potential imposed by mannitol. The mouse infection model also verified that M1PDH is essential for bacterial survival during infection. To further support the use of M1PDH as an antibacterial target, we identified dihydrocelastrol (DHCL) as a competitive inhibitor of S. aureus M1PDH (SaM1PDH) and confirmed that DHCL effectively reduces bacterial cell viability during host infection. To explain physiological functions of SaM1PDH at the atomic level, the crystal structure of SaM1PDH was determined at 1.7-Å resolution. Structure-based mutation analyses and DHCL molecular docking to the SaM1PDH active site followed by functional assay identified key residues in the active site and provided the action mechanism of DHCL. Collectively, we propose SaM1PDH as a target for antibiotic development based on its physiological roles with the goals of expanding the repertory of antibiotic targets to fight antimicrobial resistance and providing essential knowledge for developing potent inhibitors of SaM1PDH based on structure-function studies.IMPORTANCE Due to the shortage of effective antibiotics against drug-resistant Staphylococcus aureus, new targets are urgently required to develop next-generation antibiotics. We investigated mannitol-1-phosphate dehydrogenase of S. aureus USA300 (SaM1PDH), a key enzyme regulating intracellular mannitol levels, and explored the possibility of using SaM1PDH as a target for developing antibiotic. Since mannitol is necessary for maintaining the cellular redox and osmotic potential, the homeostatic imbalance caused by treatment with a SaM1PDH inhibitor or knockout of the gene encoding SaM1PDH results in bacterial cell death through oxidative and/or mannitol-dependent cytolysis. We elucidated the molecular mechanism of SaM1PDH and the structural basis of substrate and inhibitor recognition by enzymatic and structural analyses of SaM1PDH. Our results strongly support the concept that targeting of SaM1PDH represents an alternative strategy for developing a new class of antibiotics that cause bacterial cell death not by blocking key cellular machinery but by inducing cytolysis and reducing stress tolerance through inhibition of the mannitol pathway.

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Accepted/In Press date: 5 June 2019
Published date: 9 July 2019

Identifiers

Local EPrints ID: 432773
URI: http://eprints.soton.ac.uk/id/eprint/432773
ISSN: 2150-7511
PURE UUID: 1ff55bb6-3ab6-433d-837d-f80fce7fe70f
ORCID for Seung Seo Lee: ORCID iD orcid.org/0000-0002-8598-3303

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Date deposited: 26 Jul 2019 16:30
Last modified: 26 Nov 2021 03:00

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Contributors

Author: Thanh Nguyen
Author: Truc Kim
Author: Hai Minh Ta
Author: Won Sik Yeo
Author: Jongkeun Choi
Author: Pushpak Mizar
Author: Seung Seo Lee ORCID iD
Author: Taeok Bae
Author: Akhilesh Kumar Chaurasia
Author: Kyeong Kyu Kim

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