Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds
The obligate intracellular bacterium <i>Chlamydia trachomatis</i> is the major causative agent of bacterial sexually transmitted diseases worldwide. In infected cells, the ceramide transport protein (CERT) is recruited to inclusions, where <i>C. trachomatis</i> replicates usi...
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| Format: | Article |
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MDPI AG
2022-11-01
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| Series: | International Journal of Molecular Sciences |
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| Online Access: | https://www.mdpi.com/1422-0067/23/23/14697 |
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| author | Keigo Kumagai Shota Sakai Masaharu Ueno Michiyo Kataoka Shu Kobayashi Kentaro Hanada |
| author_facet | Keigo Kumagai Shota Sakai Masaharu Ueno Michiyo Kataoka Shu Kobayashi Kentaro Hanada |
| author_sort | Keigo Kumagai |
| collection | DOAJ |
| description | The obligate intracellular bacterium <i>Chlamydia trachomatis</i> is the major causative agent of bacterial sexually transmitted diseases worldwide. In infected cells, the ceramide transport protein (CERT) is recruited to inclusions, where <i>C. trachomatis</i> replicates using host-synthesized ceramide. The ceramide is converted to sphingomyelin (SM) by a chlamydial infection-dependent SM synthesis (cidSM-synthesis) pathway, which occurs even in the absence of the SM synthases (SMS)-1 and -2 of host cells. The ceramide mimetic compound (1<i>R</i>,3<i>S</i>)-HPA-12 and the nonmimetic compound E16A, both of which are potent inhibitors of CERT, repressed the proliferation of <i>C. trachomatis</i> in HeLa cells. Unexpectedly, (1<i>R</i>,3<i>R</i>)-HPA-12, a ceramide mimetic compound that lacks CERT inhibitory activity, also exhibited potent anti-chlamydial activity. Using endogenous SMS-knockout mutant HeLa cells, we revealed that (1<i>R</i>,3<i>R</i>)-HPA-12 mildly inhibited cidSM-synthesis. In addition, LC-MS analysis revealed that (1<i>R</i>,3<i>R</i>)-HPA-12 is converted to a phosphocholine-conjugated metabolite in an infection-dependent manner. Imaging analysis with a fluorescent analog of ceramide suggested that cidSM-synthesis occurs in the bacterial bodies and/or inclusions. Collectively, these results suggested that (1<i>R</i>,3<i>R</i>)-HPA-12 exerts its anti-chlamydia activity not only as an inhibitor of cidSM-synthesis, but also via putative toxic effects of its phosphocholine adduct, which is most likely produced by the cidSM-synthesis route. |
| first_indexed | 2024-03-09T17:46:55Z |
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| id | doaj.art-e43fb9b0b9a1421ba0d657696772cebc |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-09T17:46:55Z |
| publishDate | 2022-11-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-e43fb9b0b9a1421ba0d657696772cebc2023-11-24T11:06:21ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672022-11-0123231469710.3390/ijms232314697Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic CompoundsKeigo Kumagai0Shota Sakai1Masaharu Ueno2Michiyo Kataoka3Shu Kobayashi4Kentaro Hanada5Department of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, JapanDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, JapanDepartment of Natural Science, Graduate School of Advanced Technology and Science, Tokushima University, 2-1 Minami-jousanjima, Tokushima 770-8506, JapanDepartment of Pathology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, JapanDepartment of Chemistry, School of Science, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 110-0033, JapanDepartment of Biochemistry and Cell Biology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo 162-8640, JapanThe obligate intracellular bacterium <i>Chlamydia trachomatis</i> is the major causative agent of bacterial sexually transmitted diseases worldwide. In infected cells, the ceramide transport protein (CERT) is recruited to inclusions, where <i>C. trachomatis</i> replicates using host-synthesized ceramide. The ceramide is converted to sphingomyelin (SM) by a chlamydial infection-dependent SM synthesis (cidSM-synthesis) pathway, which occurs even in the absence of the SM synthases (SMS)-1 and -2 of host cells. The ceramide mimetic compound (1<i>R</i>,3<i>S</i>)-HPA-12 and the nonmimetic compound E16A, both of which are potent inhibitors of CERT, repressed the proliferation of <i>C. trachomatis</i> in HeLa cells. Unexpectedly, (1<i>R</i>,3<i>R</i>)-HPA-12, a ceramide mimetic compound that lacks CERT inhibitory activity, also exhibited potent anti-chlamydial activity. Using endogenous SMS-knockout mutant HeLa cells, we revealed that (1<i>R</i>,3<i>R</i>)-HPA-12 mildly inhibited cidSM-synthesis. In addition, LC-MS analysis revealed that (1<i>R</i>,3<i>R</i>)-HPA-12 is converted to a phosphocholine-conjugated metabolite in an infection-dependent manner. Imaging analysis with a fluorescent analog of ceramide suggested that cidSM-synthesis occurs in the bacterial bodies and/or inclusions. Collectively, these results suggested that (1<i>R</i>,3<i>R</i>)-HPA-12 exerts its anti-chlamydia activity not only as an inhibitor of cidSM-synthesis, but also via putative toxic effects of its phosphocholine adduct, which is most likely produced by the cidSM-synthesis route.https://www.mdpi.com/1422-0067/23/23/14697infectious disease<i>Chlamydia trachomatis</i>bacterial metabolismantibioticsceramidesphingolipid |
| spellingShingle | Keigo Kumagai Shota Sakai Masaharu Ueno Michiyo Kataoka Shu Kobayashi Kentaro Hanada Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds International Journal of Molecular Sciences infectious disease <i>Chlamydia trachomatis</i> bacterial metabolism antibiotics ceramide sphingolipid |
| title | Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds |
| title_full | Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds |
| title_fullStr | Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds |
| title_full_unstemmed | Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds |
| title_short | Chlamydial Infection-Dependent Synthesis of Sphingomyelin as a Novel Anti-Chlamydial Target of Ceramide Mimetic Compounds |
| title_sort | chlamydial infection dependent synthesis of sphingomyelin as a novel anti chlamydial target of ceramide mimetic compounds |
| topic | infectious disease <i>Chlamydia trachomatis</i> bacterial metabolism antibiotics ceramide sphingolipid |
| url | https://www.mdpi.com/1422-0067/23/23/14697 |
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