Publicação
Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus
| Resumo: | A multigene polysaccharide utilization locus (PUL) encoding enzymes and surface carbohydrate (glycan)-binding proteins (SGBPs) was recently identified in prominent members of Bacteroidetes in the human gut and characterized in Bacteroides ovatus. This PUL-encoded system specifically targets mixed-linkage β1,3-1,4-glucans, a group of diet-derived carbohydrates that promote a healthy microbiota and have potential as prebiotics. The BoSGBPMLG-A protein encoded by the BACOVA_2743 gene is a SusD-like protein that plays a key role in the PUL's specificity and functionality. Here, we perform a detailed analysis of the molecular determinants underlying carbohydrate binding by BoSGBPMLG-A, combining carbohydrate microarray technology with quantitative affinity studies and a high-resolution X-ray crystallography structure of the complex of BoSGBPMLG-A with a β1,3-1,4-nonasaccharide. We demonstrate its unique binding specificity toward β1,3-1,4-gluco-oligosaccharides, with increasing binding affinities up to the octasaccharide and dependency on the number and position of β1,3 linkages. The interaction is defined by a 41-Å-long extended binding site that accommodates the oligosaccharide in a mode distinct from that of previously described bacterial β1,3-1,4-glucan-binding proteins. In addition to the shape complementarity mediated by CH-π interactions, a complex hydrogen bonding network complemented by a high number of key ordered water molecules establishes additional specific interactions with the oligosaccharide. These support the twisted conformation of the β-glucan backbone imposed by the β1,3 linkages and explain the dependency on the oligosaccharide chain length. We propose that the specificity of the PUL conferred by BoSGBPMLG-A to import long β1,3-1,4-glucan oligosaccharides to the bacterial periplasm allows Bacteroidetes to outcompete bacteria that lack this PUL for utilization of β1,3-1,4-glucans. IMPORTANCE With the knowledge of bacterial gene systems encoding proteins that target dietary carbohydrates as a source of nutrients and their importance for human health, major efforts are being made to understand carbohydrate recognition by various commensal bacteria. Here, we describe an integrative strategy that combines carbohydrate microarray technology with structural studies to further elucidate the molecular determinants of carbohydrate recognition by BoSGBPMLG-A, a key protein expressed at the surface of Bacteroides ovatus for utilization of mixed-linkage β1,3-1,4-glucans. We have mapped at high resolution interactions that occur at the binding site of BoSGBPMLG-A and provide evidence for the role of key water-mediated interactions for fine specificity and affinity. Understanding at the molecular level how commensal bacteria, such as prominent members of Bacteroidetes, can differentially utilize dietary carbohydrates with potential prebiotic activities will shed light on possible ways to modulate the microbiome to promote human health. |
|---|---|
| Autores principais: | Correia, Viviana G. |
| Outros Autores: | Trovão, Filipa; Pinheiro, Benedita A.; Brás, Joana L. A.; Silva, Lisete M.; Nunes, Cláudia; Coimbra, Manuel A.; Liu, Yan; Feizi, Ten; Fontes, Carlos M. G. A.; Mulloy, Barbara; Chai, Wengang; Carvalho, Ana Luísa; Palma, Angelina S. |
| Assunto: | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| Ano: | 2021 |
| País: | Portugal |
| Tipo de documento: | artigo |
| Tipo de acesso: | acesso aberto |
| Instituição associada: | Universidade Nova de Lisboa |
| Idioma: | inglês |
| Origem: | Repositório Institucional da UNL |
| _version_ | 1868983136359022592 |
|---|---|
| author | Correia, Viviana G. |
| author2 | Trovão, Filipa Pinheiro, Benedita A. Brás, Joana L. A. Silva, Lisete M. Nunes, Cláudia Coimbra, Manuel A. Liu, Yan Feizi, Ten Fontes, Carlos M. G. A. Mulloy, Barbara Chai, Wengang Carvalho, Ana Luísa Palma, Angelina S. |
| author2_role | author author author author author author author author author author author author author |
| author_facet | Correia, Viviana G. Trovão, Filipa Pinheiro, Benedita A. Brás, Joana L. A. Silva, Lisete M. Nunes, Cláudia Coimbra, Manuel A. Liu, Yan Feizi, Ten Fontes, Carlos M. G. A. Mulloy, Barbara Chai, Wengang Carvalho, Ana Luísa Palma, Angelina S. |
| author_role | author |
| contributor_name_str_mv | UCIBIO - Applied Molecular Biosciences Unit DQ - Departamento de Química American Society for Microbiology RUN |
| country_str | PT |
| creators_json_txt | [{\"Person.name\":\"Correia, Viviana G.\"},{\"Person.name\":\"Trovão, Filipa\"},{\"Person.name\":\"Pinheiro, Benedita A.\"},{\"Person.name\":\"Brás, Joana L. A.\"},{\"Person.name\":\"Silva, Lisete M.\"},{\"Person.name\":\"Nunes, Cláudia\"},{\"Person.name\":\"Coimbra, Manuel A.\"},{\"Person.name\":\"Liu, Yan\"},{\"Person.name\":\"Feizi, Ten\"},{\"Person.name\":\"Fontes, Carlos M. G. A.\"},{\"Person.name\":\"Mulloy, Barbara\"},{\"Person.name\":\"Chai, Wengang\"},{\"Person.name\":\"Carvalho, Ana Luísa\"},{\"Person.name\":\"Palma, Angelina S.\"}] |
| datacite.contributors.contributor.contributorName.fl_str_mv | UCIBIO - Applied Molecular Biosciences Unit DQ - Departamento de Química American Society for Microbiology RUN |
| datacite.creators.creator.creatorName.fl_str_mv | Correia, Viviana G. Trovão, Filipa Pinheiro, Benedita A. Brás, Joana L. A. Silva, Lisete M. Nunes, Cláudia Coimbra, Manuel A. Liu, Yan Feizi, Ten Fontes, Carlos M. G. A. Mulloy, Barbara Chai, Wengang Carvalho, Ana Luísa Palma, Angelina S. |
| datacite.date.Accepted.fl_str_mv | 2021-11-24T00:00:00Z |
| datacite.date.available.fl_str_mv | 2021-12-02T23:47:45Z |
| datacite.date.embargoed.fl_str_mv | 2021-12-02T23:47:45Z |
| datacite.rights.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| datacite.subjects.subject.fl_str_mv | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| datacite.titles.title.fl_str_mv | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| dc.contributor.none.fl_str_mv | UCIBIO - Applied Molecular Biosciences Unit DQ - Departamento de Química American Society for Microbiology RUN |
| dc.creator.none.fl_str_mv | Correia, Viviana G. Trovão, Filipa Pinheiro, Benedita A. Brás, Joana L. A. Silva, Lisete M. Nunes, Cláudia Coimbra, Manuel A. Liu, Yan Feizi, Ten Fontes, Carlos M. G. A. Mulloy, Barbara Chai, Wengang Carvalho, Ana Luísa Palma, Angelina S. |
| dc.date.Accepted.fl_str_mv | 2021-11-24T00:00:00Z |
| dc.date.available.fl_str_mv | 2021-12-02T23:47:45Z |
| dc.date.embargoed.fl_str_mv | 2021-12-02T23:47:45Z |
| dc.format.none.fl_str_mv | application/pdf |
| dc.identifier.none.fl_str_mv | http://hdl.handle.net/10362/128612 |
| dc.language.none.fl_str_mv | eng |
| dc.rights.none.fl_str_mv | http://purl.org/coar/access_right/c_abf2 |
| dc.subject.none.fl_str_mv | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| dc.title.fl_str_mv | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| dc.type.none.fl_str_mv | http://purl.org/coar/resource_type/c_6501 |
| description | A multigene polysaccharide utilization locus (PUL) encoding enzymes and surface carbohydrate (glycan)-binding proteins (SGBPs) was recently identified in prominent members of Bacteroidetes in the human gut and characterized in Bacteroides ovatus. This PUL-encoded system specifically targets mixed-linkage β1,3-1,4-glucans, a group of diet-derived carbohydrates that promote a healthy microbiota and have potential as prebiotics. The BoSGBPMLG-A protein encoded by the BACOVA_2743 gene is a SusD-like protein that plays a key role in the PUL's specificity and functionality. Here, we perform a detailed analysis of the molecular determinants underlying carbohydrate binding by BoSGBPMLG-A, combining carbohydrate microarray technology with quantitative affinity studies and a high-resolution X-ray crystallography structure of the complex of BoSGBPMLG-A with a β1,3-1,4-nonasaccharide. We demonstrate its unique binding specificity toward β1,3-1,4-gluco-oligosaccharides, with increasing binding affinities up to the octasaccharide and dependency on the number and position of β1,3 linkages. The interaction is defined by a 41-Å-long extended binding site that accommodates the oligosaccharide in a mode distinct from that of previously described bacterial β1,3-1,4-glucan-binding proteins. In addition to the shape complementarity mediated by CH-π interactions, a complex hydrogen bonding network complemented by a high number of key ordered water molecules establishes additional specific interactions with the oligosaccharide. These support the twisted conformation of the β-glucan backbone imposed by the β1,3 linkages and explain the dependency on the oligosaccharide chain length. We propose that the specificity of the PUL conferred by BoSGBPMLG-A to import long β1,3-1,4-glucan oligosaccharides to the bacterial periplasm allows Bacteroidetes to outcompete bacteria that lack this PUL for utilization of β1,3-1,4-glucans. IMPORTANCE With the knowledge of bacterial gene systems encoding proteins that target dietary carbohydrates as a source of nutrients and their importance for human health, major efforts are being made to understand carbohydrate recognition by various commensal bacteria. Here, we describe an integrative strategy that combines carbohydrate microarray technology with structural studies to further elucidate the molecular determinants of carbohydrate recognition by BoSGBPMLG-A, a key protein expressed at the surface of Bacteroides ovatus for utilization of mixed-linkage β1,3-1,4-glucans. We have mapped at high resolution interactions that occur at the binding site of BoSGBPMLG-A and provide evidence for the role of key water-mediated interactions for fine specificity and affinity. Understanding at the molecular level how commensal bacteria, such as prominent members of Bacteroidetes, can differentially utilize dietary carbohydrates with potential prebiotic activities will shed light on possible ways to modulate the microbiome to promote human health. |
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| person_str_mv | Correia, Viviana G. Trovão, Filipa Pinheiro, Benedita A. Brás, Joana L. A. Silva, Lisete M. Nunes, Cláudia Coimbra, Manuel A. Liu, Yan Feizi, Ten Fontes, Carlos M. G. A. Mulloy, Barbara Chai, Wengang Carvalho, Ana Luísa Palma, Angelina S. |
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| spelling | engenA multigene polysaccharide utilization locus (PUL) encoding enzymes and surface carbohydrate (glycan)-binding proteins (SGBPs) was recently identified in prominent members of Bacteroidetes in the human gut and characterized in Bacteroides ovatus. This PUL-encoded system specifically targets mixed-linkage β1,3-1,4-glucans, a group of diet-derived carbohydrates that promote a healthy microbiota and have potential as prebiotics. The BoSGBPMLG-A protein encoded by the BACOVA_2743 gene is a SusD-like protein that plays a key role in the PUL's specificity and functionality. Here, we perform a detailed analysis of the molecular determinants underlying carbohydrate binding by BoSGBPMLG-A, combining carbohydrate microarray technology with quantitative affinity studies and a high-resolution X-ray crystallography structure of the complex of BoSGBPMLG-A with a β1,3-1,4-nonasaccharide. We demonstrate its unique binding specificity toward β1,3-1,4-gluco-oligosaccharides, with increasing binding affinities up to the octasaccharide and dependency on the number and position of β1,3 linkages. The interaction is defined by a 41-Å-long extended binding site that accommodates the oligosaccharide in a mode distinct from that of previously described bacterial β1,3-1,4-glucan-binding proteins. In addition to the shape complementarity mediated by CH-π interactions, a complex hydrogen bonding network complemented by a high number of key ordered water molecules establishes additional specific interactions with the oligosaccharide. These support the twisted conformation of the β-glucan backbone imposed by the β1,3 linkages and explain the dependency on the oligosaccharide chain length. We propose that the specificity of the PUL conferred by BoSGBPMLG-A to import long β1,3-1,4-glucan oligosaccharides to the bacterial periplasm allows Bacteroidetes to outcompete bacteria that lack this PUL for utilization of β1,3-1,4-glucans. IMPORTANCE With the knowledge of bacterial gene systems encoding proteins that target dietary carbohydrates as a source of nutrients and their importance for human health, major efforts are being made to understand carbohydrate recognition by various commensal bacteria. Here, we describe an integrative strategy that combines carbohydrate microarray technology with structural studies to further elucidate the molecular determinants of carbohydrate recognition by BoSGBPMLG-A, a key protein expressed at the surface of Bacteroides ovatus for utilization of mixed-linkage β1,3-1,4-glucans. We have mapped at high resolution interactions that occur at the binding site of BoSGBPMLG-A and provide evidence for the role of key water-mediated interactions for fine specificity and affinity. Understanding at the molecular level how commensal bacteria, such as prominent members of Bacteroidetes, can differentially utilize dietary carbohydrates with potential prebiotic activities will shed light on possible ways to modulate the microbiome to promote human health.application/pdfenMapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatusCorreia, Viviana G.Trovão, FilipaPinheiro, Benedita A.Brás, Joana L. A.Silva, Lisete M.Nunes, CláudiaCoimbra, Manuel A.Liu, YanFeizi, TenFontes, Carlos M. G. A.Mulloy, BarbaraChai, WengangCarvalho, Ana LuísaPalma, Angelina S.UCIBIO - Applied Molecular Biosciences UnitDQ - Departamento de QuímicaAmerican Society for MicrobiologyHostingInstitutionOrganizationalRUNe-mailmailto:run@unl.ptrun@unl.ptISSNIsPartOf2165-0497URNIsPartOfPURE: 35125874URNIsPartOfPURE UUID: 0f923c5c-f2ec-49e8-9e48-6bf8356f6993URNIsPartOfPubMed: 34817219URNIsPartOfORCID: /0000-0002-3824-0240/work/104151667URNIsPartOfORCID: /0000-0001-5797-6555/work/104151934URNIsPartOfScopus: 85122734601DOIIsPartOf10.1128/Spectrum.01826-212021-12-02T23:47:45Z2021-11-242021-11-24T00:00:00ZHandlehttp://hdl.handle.net/10362/128612http://purl.org/coar/access_right/c_abf2open accessβ-glucanBacteroides ovatucarbohydrate microarrayspolysaccharide utilization lociprotein-carbohydrate interactionsSusD-like proteinsX-ray crystallographySDG 3 - Good Health and Well-being3906593 bytesFundação para a Ciência e a TecnologiaDeciphering carbohydrate: protein interactions in the Human Microbiome - A combined High-throughput and structural approachOECrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaTranslating the human-microbiome molecular cross-talk using glycan microarray and structural biology strategiesOECrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaAssociated Laboratory for Green Chemistry - Clean Technologies and Processes6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaCICECO-Aveiro Institute of Materials6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaCICECO-Aveiro Institute of Materials6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaApplied Molecular Biosciences Unit6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871Fundação para a Ciência e a TecnologiaApplied Molecular Biosciences Unit6817 - DCRRNI IDCrossref Funder IDhttp://doi.org/10.13039/501100001871literaturehttp://purl.org/coar/resource_type/c_6501journal articlehttp://purl.org/coar/access_right/c_abf2application/pdffulltexthttps://run.unl.pt/bitstreams/7dc5a369-de46-4791-9c29-fd79366fefef/download |
| spellingShingle | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus Correia, Viviana G. β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| status | SINGLETON |
| subject.fl_str_mv | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| title | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| title_full | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| title_fullStr | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| title_full_unstemmed | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| title_short | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| title_sort | Mapping Molecular Recognition of β1,3-1,4-Glucans by a Surface Glycan-Binding Protein from the Human Gut Symbiont Bacteroides ovatus |
| topic | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| topic_facet | β-glucan Bacteroides ovatu carbohydrate microarrays polysaccharide utilization loci protein-carbohydrate interactions SusD-like proteins X-ray crystallography SDG 3 - Good Health and Well-being |
| url | http://hdl.handle.net/10362/128612 |
| visible | 1 |