The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism
Streptococcus pneumoniae, a major cause of pneumonia, sepsis, and meningitis worldwide, has the nasopharynges of small children as its main ecological niche. Depletion of pneumococci from this niche would reduce the disease burden and could be achieved using small molecules with narrow-spectrum anti...
| Main Authors: | , , , , , , , , |
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| Format: | Journal Article |
| Language: | English |
| Published: |
2021
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| Online Access: | https://hdl.handle.net/10356/146001 |
| _version_ | 1826116458405429248 |
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| author | Reithuber, Elisabeth Nannapaneni, Priyanka Rzhepishevska, Olena Lindgren, Anders E. G. Ilchenko, Oleksandr Normark, Staffan Almqvist, Fredrik Henriques-Normark, Birgitta Mellroth, Peter |
| author2 | Lee Kong Chian School of Medicine (LKCMedicine) |
| author_facet | Lee Kong Chian School of Medicine (LKCMedicine) Reithuber, Elisabeth Nannapaneni, Priyanka Rzhepishevska, Olena Lindgren, Anders E. G. Ilchenko, Oleksandr Normark, Staffan Almqvist, Fredrik Henriques-Normark, Birgitta Mellroth, Peter |
| author_sort | Reithuber, Elisabeth |
| collection | NTU |
| description | Streptococcus pneumoniae, a major cause of pneumonia, sepsis, and meningitis worldwide, has the nasopharynges of small children as its main ecological niche. Depletion of pneumococci from this niche would reduce the disease burden and could be achieved using small molecules with narrow-spectrum antibacterial activity. We identified the alkylated dicyclohexyl carboxylic acid 2CCA-1 as a potent inducer of autolysin-mediated lysis of S. pneumoniae, while having low activity against Staphylococcus aureus 2CCA-1-resistant strains were found to have inactivating mutations in fakB3, known to be required for uptake of host polyunsaturated fatty acids, as well as through inactivation of the transcriptional regulator gene fabT, vital for endogenous, de novo fatty acid synthesis regulation. Structure activity relationship exploration revealed that, besides the central dicyclohexyl group, the fatty acid-like structural features of 2CCA-1 were essential for its activity. The lysis-inducing activity of 2CCA-1 was considerably more potent than that of free fatty acids and required growing bacteria, suggesting that 2CCA-1 needs to be metabolized to exert its antimicrobial activity. Total lipid analysis of 2CCA-1 treated bacteria identified unique masses that were modeled to 2CCA-1 containing lysophosphatidic and phosphatidic acid in wild-type but not in fakB3 mutant bacteria. This suggests that 2CCA-1 is metabolized as a fatty acid via FakB3 and utilized as a phospholipid building block, leading to accumulation of toxic phospholipid species. Analysis of FabT-mediated fakB3 expression elucidates how the pneumococcus could ensure membrane homeostasis and concurrent economic use of host-derived fatty acids.IMPORTANCE Fatty acid biosynthesis is an attractive antibiotic target, as it affects the supply of membrane phospholipid building blocks. In Streptococcus pneumoniae, it is not sufficient to target only the endogenous fatty acid synthesis machinery, as uptake of host fatty acids may bypass this inhibition. Here, we describe a small-molecule compound, 2CCA-1, with potent bactericidal activity that upon interactions with the fatty acid binding protein FakB3, which is present in a limited number of Gram-positive species, becomes metabolized and incorporated as a toxic phospholipid species. Resistance to 2CCA-1 developed specifically in fakB3 and the regulatory gene fabT These mutants reveal a regulatory connection between the extracellular polyunsaturated fatty acid metabolism and endogenous fatty acid synthesis in S. pneumoniae, which could ensure balance between efficient scavenging of host polyunsaturated fatty acids and membrane homeostasis. The data might be useful in the identification of narrow-spectrum treatment strategies to selectively target members of the Lactobacillales such as S. pneumoniae. |
| first_indexed | 2024-10-01T04:11:43Z |
| format | Journal Article |
| id | ntu-10356/146001 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2024-10-01T04:11:43Z |
| publishDate | 2021 |
| record_format | dspace |
| spelling | ntu-10356/1460012021-01-23T20:11:47Z The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism Reithuber, Elisabeth Nannapaneni, Priyanka Rzhepishevska, Olena Lindgren, Anders E. G. Ilchenko, Oleksandr Normark, Staffan Almqvist, Fredrik Henriques-Normark, Birgitta Mellroth, Peter Lee Kong Chian School of Medicine (LKCMedicine) Singapore Centre for Environmental Life Sciences and Engineering Science::Medicine Streptococcus Pneumoniae Pneumococci Streptococcus pneumoniae, a major cause of pneumonia, sepsis, and meningitis worldwide, has the nasopharynges of small children as its main ecological niche. Depletion of pneumococci from this niche would reduce the disease burden and could be achieved using small molecules with narrow-spectrum antibacterial activity. We identified the alkylated dicyclohexyl carboxylic acid 2CCA-1 as a potent inducer of autolysin-mediated lysis of S. pneumoniae, while having low activity against Staphylococcus aureus 2CCA-1-resistant strains were found to have inactivating mutations in fakB3, known to be required for uptake of host polyunsaturated fatty acids, as well as through inactivation of the transcriptional regulator gene fabT, vital for endogenous, de novo fatty acid synthesis regulation. Structure activity relationship exploration revealed that, besides the central dicyclohexyl group, the fatty acid-like structural features of 2CCA-1 were essential for its activity. The lysis-inducing activity of 2CCA-1 was considerably more potent than that of free fatty acids and required growing bacteria, suggesting that 2CCA-1 needs to be metabolized to exert its antimicrobial activity. Total lipid analysis of 2CCA-1 treated bacteria identified unique masses that were modeled to 2CCA-1 containing lysophosphatidic and phosphatidic acid in wild-type but not in fakB3 mutant bacteria. This suggests that 2CCA-1 is metabolized as a fatty acid via FakB3 and utilized as a phospholipid building block, leading to accumulation of toxic phospholipid species. Analysis of FabT-mediated fakB3 expression elucidates how the pneumococcus could ensure membrane homeostasis and concurrent economic use of host-derived fatty acids.IMPORTANCE Fatty acid biosynthesis is an attractive antibiotic target, as it affects the supply of membrane phospholipid building blocks. In Streptococcus pneumoniae, it is not sufficient to target only the endogenous fatty acid synthesis machinery, as uptake of host fatty acids may bypass this inhibition. Here, we describe a small-molecule compound, 2CCA-1, with potent bactericidal activity that upon interactions with the fatty acid binding protein FakB3, which is present in a limited number of Gram-positive species, becomes metabolized and incorporated as a toxic phospholipid species. Resistance to 2CCA-1 developed specifically in fakB3 and the regulatory gene fabT These mutants reveal a regulatory connection between the extracellular polyunsaturated fatty acid metabolism and endogenous fatty acid synthesis in S. pneumoniae, which could ensure balance between efficient scavenging of host polyunsaturated fatty acids and membrane homeostasis. The data might be useful in the identification of narrow-spectrum treatment strategies to selectively target members of the Lactobacillales such as S. pneumoniae. Published version 2021-01-20T03:50:32Z 2021-01-20T03:50:32Z 2020 Journal Article Reithuber, E., Nannapaneni, P., Rzhepishevska, O., Lindgren, A. E. G., Ilchenko, O., Normark, S., . . . Mellroth, P. (2020). The Bactericidal Fatty Acid Mimetic 2CCA-1 Selectively Targets Pneumococcal Extracellular Polyunsaturated Fatty Acid Metabolism. MBio, 11(6), e03027-20-. doi:10.1128/mbio.03027-20 2150-7511 https://hdl.handle.net/10356/146001 10.1128/mBio.03027-20 33323510 2-s2.0-85098532621 6 11 en mBio © 2020 Reithuber et al. This is anMopen-access article distributed under the terms of the Creative Commons Attribution 4.0 International license. application/pdf |
| spellingShingle | Science::Medicine Streptococcus Pneumoniae Pneumococci Reithuber, Elisabeth Nannapaneni, Priyanka Rzhepishevska, Olena Lindgren, Anders E. G. Ilchenko, Oleksandr Normark, Staffan Almqvist, Fredrik Henriques-Normark, Birgitta Mellroth, Peter The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title | The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title_full | The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title_fullStr | The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title_full_unstemmed | The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title_short | The bactericidal fatty acid mimetic 2CCA-1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| title_sort | bactericidal fatty acid mimetic 2cca 1 selectively targets pneumococcal extracellular polyunsaturated fatty acid metabolism |
| topic | Science::Medicine Streptococcus Pneumoniae Pneumococci |
| url | https://hdl.handle.net/10356/146001 |
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