Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci
ABSTRACT Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and no effective vaccine exists currently. In this study, the structure, biological properties, and vaccine potential of the Ng-adhesin complex protein (Ng-ACP) are presented. T...
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American Society for Microbiology
2018
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oai:doaj.org-article:e2941d24a855434e9c37e27add646cd82021-11-15T15:22:26ZStructure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci10.1128/mSphere.00331-182379-5042https://doaj.org/article/e2941d24a855434e9c37e27add646cd82018-10-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00331-18https://doaj.org/toc/2379-5042ABSTRACT Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and no effective vaccine exists currently. In this study, the structure, biological properties, and vaccine potential of the Ng-adhesin complex protein (Ng-ACP) are presented. The crystal structure of recombinant Ng-ACP (rNg-ACP) protein was solved at 1.65 Å. Diversity and conservation of Ng-ACP were examined in different Neisseria species and gonococcal isolates (https://pubmlst.org/neisseria/ database) in silico, and protein expression among 50 gonococcal strains in the Centers for Disease Control and Prevention/Food and Drug Administration (CDCP/FDA) AR Isolate Bank was examined by Western blotting. Murine antisera were raised to allele 10 (strain P9-17)-encoded rNg-ACP protein with different adjuvants and examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and a human serum bactericidal assay. Rabbit antiserum to rNg-ACP was tested for its ability to prevent Ng-ACP from inhibiting human lysozyme activity in vitro. Ng-ACP is structurally homologous to Neisseria meningitidis ACP and MliC/PliC lysozyme inhibitors. Gonococci expressed predominantly allele 10- and allele 6-encoded Ng-ACP (81% and 15% of isolates, respectively). Murine antisera were bactericidal (titers of 64 to 512, P < 0.05) for the homologous P9-17 strain and heterologous (allele 6) FA1090 strain. Rabbit anti-rNg-ACP serum prevented Ng-ACP from inhibiting human lysozyme with ∼100% efficiency. Ng-ACP protein was expressed by all 50 gonococcal isolates examined with minor differences in the relative levels of expression. rNg-ACP is a potential vaccine candidate that induces antibodies that (i) are bactericidal and (ii) prevent the gonococcus from inhibiting the lytic activity of an innate defense molecule. IMPORTANCE Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and the organism is listed by the World Health Organization as a high-priority pathogen for research and development of new control measures, including vaccines. In this study, we demonstrated that the N. gonorrhoeae adhesin complex protein (Ng-ACP) was conserved and expressed by 50 gonococcal strains and that recombinant proteins induced antibodies in mice that killed the bacteria in vitro. We determined the structure of Ng-ACP by X-ray crystallography and investigated structural conservation with Neisseria meningitidis ACP and MliC/PliC proteins from other bacteria which act as inhibitors of the human innate defense molecule lysozyme. These findings are important and suggest that Ng-ACP could provide a potential dual target for tackling gonococcal infections.Hannia Liliana Almonacid-MendozaMaría Victoria HumbertAiste DijokaiteDavid W. ClearyYiwen SooMiao-Chiu HungChristian M. OrrMoritz M. MachelettIvo TewsMyron ChristodoulidesAmerican Society for MicrobiologyarticleNGO1981Neisseria gonorrhoeaeadhesin complex proteinbactericidal antibodycrystal structurerecombinant protein productionMicrobiologyQR1-502ENmSphere, Vol 3, Iss 5 (2018) |
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| topic |
NGO1981 Neisseria gonorrhoeae adhesin complex protein bactericidal antibody crystal structure recombinant protein production Microbiology QR1-502 |
| spellingShingle |
NGO1981 Neisseria gonorrhoeae adhesin complex protein bactericidal antibody crystal structure recombinant protein production Microbiology QR1-502 Hannia Liliana Almonacid-Mendoza María Victoria Humbert Aiste Dijokaite David W. Cleary Yiwen Soo Miao-Chiu Hung Christian M. Orr Moritz M. Machelett Ivo Tews Myron Christodoulides Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| description |
ABSTRACT Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and no effective vaccine exists currently. In this study, the structure, biological properties, and vaccine potential of the Ng-adhesin complex protein (Ng-ACP) are presented. The crystal structure of recombinant Ng-ACP (rNg-ACP) protein was solved at 1.65 Å. Diversity and conservation of Ng-ACP were examined in different Neisseria species and gonococcal isolates (https://pubmlst.org/neisseria/ database) in silico, and protein expression among 50 gonococcal strains in the Centers for Disease Control and Prevention/Food and Drug Administration (CDCP/FDA) AR Isolate Bank was examined by Western blotting. Murine antisera were raised to allele 10 (strain P9-17)-encoded rNg-ACP protein with different adjuvants and examined by enzyme-linked immunosorbent assay (ELISA), Western blotting, and a human serum bactericidal assay. Rabbit antiserum to rNg-ACP was tested for its ability to prevent Ng-ACP from inhibiting human lysozyme activity in vitro. Ng-ACP is structurally homologous to Neisseria meningitidis ACP and MliC/PliC lysozyme inhibitors. Gonococci expressed predominantly allele 10- and allele 6-encoded Ng-ACP (81% and 15% of isolates, respectively). Murine antisera were bactericidal (titers of 64 to 512, P < 0.05) for the homologous P9-17 strain and heterologous (allele 6) FA1090 strain. Rabbit anti-rNg-ACP serum prevented Ng-ACP from inhibiting human lysozyme with ∼100% efficiency. Ng-ACP protein was expressed by all 50 gonococcal isolates examined with minor differences in the relative levels of expression. rNg-ACP is a potential vaccine candidate that induces antibodies that (i) are bactericidal and (ii) prevent the gonococcus from inhibiting the lytic activity of an innate defense molecule. IMPORTANCE Neisseria gonorrhoeae (gonococcus [Ng]) is the causative organism of the sexually transmitted disease gonorrhoea, and the organism is listed by the World Health Organization as a high-priority pathogen for research and development of new control measures, including vaccines. In this study, we demonstrated that the N. gonorrhoeae adhesin complex protein (Ng-ACP) was conserved and expressed by 50 gonococcal strains and that recombinant proteins induced antibodies in mice that killed the bacteria in vitro. We determined the structure of Ng-ACP by X-ray crystallography and investigated structural conservation with Neisseria meningitidis ACP and MliC/PliC proteins from other bacteria which act as inhibitors of the human innate defense molecule lysozyme. These findings are important and suggest that Ng-ACP could provide a potential dual target for tackling gonococcal infections. |
| format |
article |
| author |
Hannia Liliana Almonacid-Mendoza María Victoria Humbert Aiste Dijokaite David W. Cleary Yiwen Soo Miao-Chiu Hung Christian M. Orr Moritz M. Machelett Ivo Tews Myron Christodoulides |
| author_facet |
Hannia Liliana Almonacid-Mendoza María Victoria Humbert Aiste Dijokaite David W. Cleary Yiwen Soo Miao-Chiu Hung Christian M. Orr Moritz M. Machelett Ivo Tews Myron Christodoulides |
| author_sort |
Hannia Liliana Almonacid-Mendoza |
| title |
Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| title_short |
Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| title_full |
Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| title_fullStr |
Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| title_full_unstemmed |
Structure of the Recombinant <italic toggle="yes">Neisseria gonorrhoeae</italic> Adhesin Complex Protein (rNg-ACP) and Generation of Murine Antibodies with Bactericidal Activity against Gonococci |
| title_sort |
structure of the recombinant <italic toggle="yes">neisseria gonorrhoeae</italic> adhesin complex protein (rng-acp) and generation of murine antibodies with bactericidal activity against gonococci |
| publisher |
American Society for Microbiology |
| publishDate |
2018 |
| url |
https://doaj.org/article/e2941d24a855434e9c37e27add646cd8 |
| work_keys_str_mv |
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1718428041512747008 |