Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>
ABSTRACT Neisseria gonorrhoeae, responsible for the sexually transmitted infection gonorrhea, is an obligate human pathogen exquisitely adapted for survival on mucosal surfaces of humans. This host-pathogen relationship has resulted in evolution by N. gonorrhoeae of pathways that enable the use of h...
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| Format: | Article |
| Language: | English |
| Published: |
American Society for Microbiology
2020-06-01
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| Series: | mBio |
| Subjects: | |
| Online Access: | https://journals.asm.org/doi/10.1128/mBio.00949-20 |
| _version_ | 1818825913806094336 |
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| author | Michael T. Kammerman Aloke Bera Runrun Wu Simone A. Harrison C. Noel Maxwell Karl Lundquist Nicholas Noinaj Walter J. Chazin Cynthia Nau Cornelissen |
| author_facet | Michael T. Kammerman Aloke Bera Runrun Wu Simone A. Harrison C. Noel Maxwell Karl Lundquist Nicholas Noinaj Walter J. Chazin Cynthia Nau Cornelissen |
| author_sort | Michael T. Kammerman |
| collection | DOAJ |
| description | ABSTRACT Neisseria gonorrhoeae, responsible for the sexually transmitted infection gonorrhea, is an obligate human pathogen exquisitely adapted for survival on mucosal surfaces of humans. This host-pathogen relationship has resulted in evolution by N. gonorrhoeae of pathways that enable the use of host metalloproteins as required nutrients through the deployment of outer membrane-bound TonB-dependent transporters (TdTs). Recently, a TdT called TdfH was implicated in binding to calprotectin (CP) and in removal of the bound zinc (Zn), enabling gonococcal growth. TdfH is highly conserved among the pathogenic Neisseria species, making it a potentially promising candidate for inclusion into a gonococcal vaccine. Currently, the nature and specificity of the TdfH-CP interaction have not been determined. In this study, we found that TdfH specifically interacted with human calprotectin (hCP) and that growth of the gonococcus was supported in a TdfH-dependent manner only when hCP was available as a sole zinc source and not when mouse CP was provided. The binding interactions between TdfH and hCP were assessed using isothermal titration calorimetry where we observed a multistate model having both high-affinity and low-affinity sites of interaction. hCP has two Zn binding sites, and gonococcal growth assays using hCP mutants deficient in one or both of the Zn binding sites revealed that TdfH exhibited a site preference during Zn piracy and utilization. This report provides the first insights into the molecular mechanism of Zn piracy by neisserial TdfH and further highlights the obligate human nature of N. gonorrhoeae and the high-affinity interactions occurring between TdTs and their human ligands during pathogenesis. IMPORTANCE The dramatic rise in antimicrobial resistance among Neisseria gonorrhoeae isolates over the last few decades, paired with dwindling treatment options and the lack of a protective vaccine, has prompted increased interest in identifying new bacterial targets for the treatment and, ideally, prevention of gonococcal disease. TonB-dependent transporters are a conserved set of proteins that serve crucial functions for bacterial survival within the host. In this study, binding between the gonococcal transporter, TdfH, and calprotectin was determined to be of high affinity and host restricted. The current study identified a preferential TdfH interaction at the calprotectin dimer interface. An antigonococcal therapeutic could potentially block this site on calprotectin, interrupting Zn uptake by N. gonorrhoeae and thereby prohibiting continued bacterial growth. We describe protein-protein interactions between TdfH and calprotectin, and our findings provide the building blocks for future therapeutic or prophylactic targets. |
| first_indexed | 2024-12-19T00:19:19Z |
| format | Article |
| id | doaj.art-e6143808721949f8898784c762798033 |
| institution | Directory Open Access Journal |
| issn | 2150-7511 |
| language | English |
| last_indexed | 2024-12-19T00:19:19Z |
| publishDate | 2020-06-01 |
| publisher | American Society for Microbiology |
| record_format | Article |
| series | mBio |
| spelling | doaj.art-e6143808721949f8898784c7627980332022-12-21T20:45:37ZengAmerican Society for MicrobiologymBio2150-75112020-06-0111310.1128/mBio.00949-20Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content>Michael T. Kammerman0Aloke Bera1Runrun Wu2Simone A. Harrison3C. Noel Maxwell4Karl Lundquist5Nicholas Noinaj6Walter J. Chazin7Cynthia Nau Cornelissen8Center for Translational Immunology, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USAMarkey Center for Structural Biology, Department of Biological Science, Purdue University, West Lafayette, Indiana, USAMarkey Center for Structural Biology, Department of Biological Science, Purdue University, West Lafayette, Indiana, USADepartment of Biochemistry, Vanderbilt University, Nashville, Tennessee, USADepartment of Biochemistry, Vanderbilt University, Nashville, Tennessee, USAMarkey Center for Structural Biology, Department of Biological Science, Purdue University, West Lafayette, Indiana, USAMarkey Center for Structural Biology, Department of Biological Science, Purdue University, West Lafayette, Indiana, USADepartment of Biochemistry, Vanderbilt University, Nashville, Tennessee, USACenter for Translational Immunology, Institute for Biomedical Sciences, Georgia State University, Atlanta, Georgia, USAABSTRACT Neisseria gonorrhoeae, responsible for the sexually transmitted infection gonorrhea, is an obligate human pathogen exquisitely adapted for survival on mucosal surfaces of humans. This host-pathogen relationship has resulted in evolution by N. gonorrhoeae of pathways that enable the use of host metalloproteins as required nutrients through the deployment of outer membrane-bound TonB-dependent transporters (TdTs). Recently, a TdT called TdfH was implicated in binding to calprotectin (CP) and in removal of the bound zinc (Zn), enabling gonococcal growth. TdfH is highly conserved among the pathogenic Neisseria species, making it a potentially promising candidate for inclusion into a gonococcal vaccine. Currently, the nature and specificity of the TdfH-CP interaction have not been determined. In this study, we found that TdfH specifically interacted with human calprotectin (hCP) and that growth of the gonococcus was supported in a TdfH-dependent manner only when hCP was available as a sole zinc source and not when mouse CP was provided. The binding interactions between TdfH and hCP were assessed using isothermal titration calorimetry where we observed a multistate model having both high-affinity and low-affinity sites of interaction. hCP has two Zn binding sites, and gonococcal growth assays using hCP mutants deficient in one or both of the Zn binding sites revealed that TdfH exhibited a site preference during Zn piracy and utilization. This report provides the first insights into the molecular mechanism of Zn piracy by neisserial TdfH and further highlights the obligate human nature of N. gonorrhoeae and the high-affinity interactions occurring between TdTs and their human ligands during pathogenesis. IMPORTANCE The dramatic rise in antimicrobial resistance among Neisseria gonorrhoeae isolates over the last few decades, paired with dwindling treatment options and the lack of a protective vaccine, has prompted increased interest in identifying new bacterial targets for the treatment and, ideally, prevention of gonococcal disease. TonB-dependent transporters are a conserved set of proteins that serve crucial functions for bacterial survival within the host. In this study, binding between the gonococcal transporter, TdfH, and calprotectin was determined to be of high affinity and host restricted. The current study identified a preferential TdfH interaction at the calprotectin dimer interface. An antigonococcal therapeutic could potentially block this site on calprotectin, interrupting Zn uptake by N. gonorrhoeae and thereby prohibiting continued bacterial growth. We describe protein-protein interactions between TdfH and calprotectin, and our findings provide the building blocks for future therapeutic or prophylactic targets.https://journals.asm.org/doi/10.1128/mBio.00949-20isothermal calorimetrycalprotectinprotein-protein interactionsN. gonorrhoeaeNeisseria gonorrhoeaeTonB-dependent transporter |
| spellingShingle | Michael T. Kammerman Aloke Bera Runrun Wu Simone A. Harrison C. Noel Maxwell Karl Lundquist Nicholas Noinaj Walter J. Chazin Cynthia Nau Cornelissen Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> mBio isothermal calorimetry calprotectin protein-protein interactions N. gonorrhoeae Neisseria gonorrhoeae TonB-dependent transporter |
| title | Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> |
| title_full | Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> |
| title_fullStr | Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> |
| title_full_unstemmed | Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> |
| title_short | Molecular Insight into TdfH-Mediated Zinc Piracy from Human Calprotectin by <named-content content-type="genus-species">Neisseria gonorrhoeae</named-content> |
| title_sort | molecular insight into tdfh mediated zinc piracy from human calprotectin by named content content type genus species neisseria gonorrhoeae named content |
| topic | isothermal calorimetry calprotectin protein-protein interactions N. gonorrhoeae Neisseria gonorrhoeae TonB-dependent transporter |
| url | https://journals.asm.org/doi/10.1128/mBio.00949-20 |
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