The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases
Abstract Corynebacterium diphtheriae clade species secrete single-domain endo-β-N-acetylglucosaminidases (ENGases) that specifically bind to human IgG antibodies and hydrolyze their N297-linked glycans. Here, we define the molecular mechanisms of IgG-specific deglycosylation for the entire family of...
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Nature Portfolio
2025-07-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-025-60986-w |
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| author | Diego E. Sastre Stylianos Bournazos Maros Huliciak Barbara Ann C. Grace E. Josephine Boder Jonathan Du Nazneen Sultana Tala Azzam Trenton J. Brown Maria W. Flowers Pete Lollar Ting Xu Tatiana A. Chernova Alasdair D. Keith Meredith Keen Abigail Saltzman Ana Martinez Gascueña Beatriz Trastoy Marcelo E. Guerin Filipp Frank Eric A. Ortlund Jeffrey V. Ravetch Eric J. Sundberg |
| author_facet | Diego E. Sastre Stylianos Bournazos Maros Huliciak Barbara Ann C. Grace E. Josephine Boder Jonathan Du Nazneen Sultana Tala Azzam Trenton J. Brown Maria W. Flowers Pete Lollar Ting Xu Tatiana A. Chernova Alasdair D. Keith Meredith Keen Abigail Saltzman Ana Martinez Gascueña Beatriz Trastoy Marcelo E. Guerin Filipp Frank Eric A. Ortlund Jeffrey V. Ravetch Eric J. Sundberg |
| author_sort | Diego E. Sastre |
| collection | DOAJ |
| description | Abstract Corynebacterium diphtheriae clade species secrete single-domain endo-β-N-acetylglucosaminidases (ENGases) that specifically bind to human IgG antibodies and hydrolyze their N297-linked glycans. Here, we define the molecular mechanisms of IgG-specific deglycosylation for the entire family of corynebacterial IgG-specific ENGases, including but not limited to CU43 and CM49. By solving the crystal structure of CU43 in a 1:1 complex with the IgG1 Fc region, combined with targeted and saturation mutagenesis analysis and activity measurements using engineered antibodies, we establish an inter-protomeric mechanism of recognition and deglycosylation of IgG antibodies. Using in silico modeling, small-angle X-ray scattering and saturation mutagenesis we determine that CM49 uses a unique binding site on the Fc region, to process N297-linked glycans. Moreover, we demonstrate that CU43 treatment is highly effective in abrogating Fc effector functions in humanized mouse models, while preserving the neutralizing capacity of anti-influenza IgG antibodies, thereby conferring protection against lethal influenza challenge. |
| format | Article |
| id | doaj-art-c626d676dd6c4f10ace5c3cd75dc3e46 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | Nature Portfolio |
| record_format | Article |
| series | Nature Communications |
| spelling | doaj-art-c626d676dd6c4f10ace5c3cd75dc3e462025-08-20T03:45:34ZengNature PortfolioNature Communications2041-17232025-07-0116111710.1038/s41467-025-60986-wThe mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidasesDiego E. Sastre0Stylianos Bournazos1Maros Huliciak2Barbara Ann C. Grace3E. Josephine Boder4Jonathan Du5Nazneen Sultana6Tala Azzam7Trenton J. Brown8Maria W. Flowers9Pete Lollar10Ting Xu11Tatiana A. Chernova12Alasdair D. Keith13Meredith Keen14Abigail Saltzman15Ana Martinez Gascueña16Beatriz Trastoy17Marcelo E. Guerin18Filipp Frank19Eric A. Ortlund20Jeffrey V. Ravetch21Eric J. Sundberg22Department of Biochemistry, Emory University School of MedicineLaboratory of Molecular Genetics and Immunology, The Rockefeller UniversityDepartment of Biochemistry, Emory University School of MedicineLaboratory of Molecular Genetics and Immunology, The Rockefeller UniversityLaboratory of Molecular Genetics and Immunology, The Rockefeller UniversityDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Pediatrics, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineStructural Glycoinmunology Laboratory, Biobizkaia Health Research InstituteStructural Glycoinmunology Laboratory, Biobizkaia Health Research InstituteStructural Glycobiology Laboratory, Department of Structural and Molecular Biology; Molecular Biology Institute of Barcelona (IBMB)Department of Biochemistry, Emory University School of MedicineDepartment of Biochemistry, Emory University School of MedicineLaboratory of Molecular Genetics and Immunology, The Rockefeller UniversityDepartment of Biochemistry, Emory University School of MedicineAbstract Corynebacterium diphtheriae clade species secrete single-domain endo-β-N-acetylglucosaminidases (ENGases) that specifically bind to human IgG antibodies and hydrolyze their N297-linked glycans. Here, we define the molecular mechanisms of IgG-specific deglycosylation for the entire family of corynebacterial IgG-specific ENGases, including but not limited to CU43 and CM49. By solving the crystal structure of CU43 in a 1:1 complex with the IgG1 Fc region, combined with targeted and saturation mutagenesis analysis and activity measurements using engineered antibodies, we establish an inter-protomeric mechanism of recognition and deglycosylation of IgG antibodies. Using in silico modeling, small-angle X-ray scattering and saturation mutagenesis we determine that CM49 uses a unique binding site on the Fc region, to process N297-linked glycans. Moreover, we demonstrate that CU43 treatment is highly effective in abrogating Fc effector functions in humanized mouse models, while preserving the neutralizing capacity of anti-influenza IgG antibodies, thereby conferring protection against lethal influenza challenge.https://doi.org/10.1038/s41467-025-60986-w |
| spellingShingle | Diego E. Sastre Stylianos Bournazos Maros Huliciak Barbara Ann C. Grace E. Josephine Boder Jonathan Du Nazneen Sultana Tala Azzam Trenton J. Brown Maria W. Flowers Pete Lollar Ting Xu Tatiana A. Chernova Alasdair D. Keith Meredith Keen Abigail Saltzman Ana Martinez Gascueña Beatriz Trastoy Marcelo E. Guerin Filipp Frank Eric A. Ortlund Jeffrey V. Ravetch Eric J. Sundberg The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases Nature Communications |
| title | The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases |
| title_full | The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases |
| title_fullStr | The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases |
| title_full_unstemmed | The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases |
| title_short | The mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial IgG-specific endoglycosidases |
| title_sort | mechanistic basis for interprotomer deglycosylation of antibodies by corynebacterial igg specific endoglycosidases |
| url | https://doi.org/10.1038/s41467-025-60986-w |
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