NagS is a novel GlcNAc-6P dehydratase.
<p><b>(a)</b> Sensitivity of <i>S. coelicolor</i> mutants to GlcNAc. Spores (5 × 10<sup>5</sup> CFU) of <i>S. coelicolor</i> M145 and its mutant derivatives ∆<i>nagB</i>, SMA11, ∆<i>nagB</i>∆<i>nagS</i>, ∆<i...
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2025
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Etiketa erantsi
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| _version_ | 1849927625803300864 |
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| author | Chao Li (145513) |
| author2 | Mia Urem (22683580) Ioli Kotsogianni (9930187) Josephine Lau (20442343) Chao Du (288036) Somayah S. Elsayed (9192571) Nathaniel I. Martin (847378) Iain W. McNae (840777) Patrick Voskamp (2311429) Christoph Mayer (57204) Sébastien Rigali (724082) Navraj Pannu (10163763) Jan Pieter Abrahams (1429531) Lennart Schada von Borzyskowski (22683583) Gilles P. van Wezel (7838948) |
| author2_role | author author author author author author author author author author author author author author |
| author_facet | Chao Li (145513) Mia Urem (22683580) Ioli Kotsogianni (9930187) Josephine Lau (20442343) Chao Du (288036) Somayah S. Elsayed (9192571) Nathaniel I. Martin (847378) Iain W. McNae (840777) Patrick Voskamp (2311429) Christoph Mayer (57204) Sébastien Rigali (724082) Navraj Pannu (10163763) Jan Pieter Abrahams (1429531) Lennart Schada von Borzyskowski (22683583) Gilles P. van Wezel (7838948) |
| author_role | author |
| dc.creator.none.fl_str_mv | Chao Li (145513) Mia Urem (22683580) Ioli Kotsogianni (9930187) Josephine Lau (20442343) Chao Du (288036) Somayah S. Elsayed (9192571) Nathaniel I. Martin (847378) Iain W. McNae (840777) Patrick Voskamp (2311429) Christoph Mayer (57204) Sébastien Rigali (724082) Navraj Pannu (10163763) Jan Pieter Abrahams (1429531) Lennart Schada von Borzyskowski (22683583) Gilles P. van Wezel (7838948) |
| dc.date.none.fl_str_mv | 2025-11-25T19:00:32Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pbio.3003514.g001 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/figure/NagS_is_a_novel_GlcNAc-6P_dehydratase_/30715199 |
| dc.rights.none.fl_str_mv | CC BY 4.0 info:eu-repo/semantics/openAccess |
| dc.subject.none.fl_str_mv | Biochemistry Microbiology Ecology Developmental Biology Inorganic Chemistry Biological Sciences not elsewhere classified controls nutrient signaling also revealed 6 highlighted key residues glcnac sensing requires toxicity pathway dependent 6p deacetylase naga novel glcnac 6 key role substrate glcnac novel enzyme work uncovers wall leads unprecedented reaction substrate mycelium structural analogue streptomycetaceae </ streptomyces </ revolves around promiscuous activity nature ’ n </ multicellular lifestyle metabolic checkpoint medicine makers lytic dismantling landmark event hyphal cell highly conserved growth media function analysis clinical antibiotics central metabolism catalytic inhibitor antibiotic production active site >- acetylglucosamine 6p dehydratase |
| dc.title.none.fl_str_mv | NagS is a novel GlcNAc-6P dehydratase. |
| dc.type.none.fl_str_mv | Image Figure info:eu-repo/semantics/publishedVersion image |
| description | <p><b>(a)</b> Sensitivity of <i>S. coelicolor</i> mutants to GlcNAc. Spores (5 × 10<sup>5</sup> CFU) of <i>S. coelicolor</i> M145 and its mutant derivatives ∆<i>nagB</i>, SMA11, ∆<i>nagB</i>∆<i>nagS</i>, ∆<i>nagB</i>∆<i>nagS</i><sup>C</sup> (∆<i>nagB</i>∆<i>nagS</i> expressing <i>nagS</i>) and ∆<i>nagB</i>∆<i>nagS</i><sup>E</sup> (∆<i>nagB</i>∆<i>nagS</i> with empty plasmid pSET152) were streaked on MM agar plates with 1% mannitol (Mann) and 1% mannitol plus 10 mM GlcNAc (GlcNAc). Strains were cultured for 72 h at 30 °C. Note that <i>nagB</i> mutants can grow in the presence of 10 mM GlcNAc only when SCO4393 (<i>nagS)</i> has been mutated (suppressor SMA11) or deleted. <b>(b)</b> NagS and its role in GlcNAc sensing. Spores of M145 and ∆<i>nagS</i> were plated on MM and R5 with 0, 0.001, 0.01, 0.1, 1, 5, 10, 20, 50, 100, 150, and 200 mM GlcNAc. Note that <i>nagS</i> mutants hardly respond to GlcNAc. <b>(c)</b> Phylogenetic tree of several different types of sugar isomerase (SIS) domain enzymes in bacteria, including NagS, Glucose-6P isomerase (Pgi), Phosphoheptose isomerase (GmhA), <i>N</i>-acetylmuramic acid-6-phosphate etherase (MurQ), Glutamine-fructose-6-phosphate aminotransferase (GlmS), and putative D-galactosamine-6-phosphate deaminase (Agas). The phylogenetic tree was made by MEGA11 (Neighbour-joining method) and built based on the alignment of the amino acid sequences. <b>(d)</b> ITC analysis of NagS with <i>N</i>-acetyl-6-phosphate amino sugar metabolites, namely GlcNAc-6P, ManNAc-6P, and GalNAc-6P. Both GlcNAc-6P and ManNAc-6P are bound well by NagS, while the enzyme did not bind to GalNAc-6P. (<b>e and f</b>) confirmation of NagS products by LC-MS. Extracted ion chromatograms for GlcNAc-6-P (black trace) and its dehydrated product (pink trace) in the enzymatic reaction mixture of GlcNAc-6-P with either active NagS (e) or heat-inactivated NagS <b>(f)</b>. Peaks relating to the product (<i>m</i>/<i>z</i> = 282.0387) are indicated by red arrows. <b>(g)</b> Kinetics of NagS in the presence of 6-PG (0, 0.33, 1 mM). The <i>V</i><sub>0</sub> data used in d were plotted against the substrate concentration, and each assay was performed in triplicate and expressed as a mean ± standard error. The data underlying this Figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003514#pbio.3003514.s022" target="_blank">S1 Data</a>.</p> |
| eu_rights_str_mv | openAccess |
| id | Manara_0a8669a0f6d3e7da187ff5ff91ecc4f8 |
| identifier_str_mv | 10.1371/journal.pbio.3003514.g001 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30715199 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | NagS is a novel GlcNAc-6P dehydratase.Chao Li (145513)Mia Urem (22683580)Ioli Kotsogianni (9930187)Josephine Lau (20442343)Chao Du (288036)Somayah S. Elsayed (9192571)Nathaniel I. Martin (847378)Iain W. McNae (840777)Patrick Voskamp (2311429)Christoph Mayer (57204)Sébastien Rigali (724082)Navraj Pannu (10163763)Jan Pieter Abrahams (1429531)Lennart Schada von Borzyskowski (22683583)Gilles P. van Wezel (7838948)BiochemistryMicrobiologyEcologyDevelopmental BiologyInorganic ChemistryBiological Sciences not elsewhere classifiedcontrols nutrient signalingalso revealed 6highlighted key residuesglcnac sensing requirestoxicity pathway dependent6p deacetylase naganovel glcnac 6key rolesubstrate glcnacnovel enzymework uncoverswall leadsunprecedented reactionsubstrate myceliumstructural analoguestreptomycetaceae </streptomyces </revolves aroundpromiscuous activitynature ’n </multicellular lifestylemetabolic checkpointmedicine makerslytic dismantlinglandmark eventhyphal cellhighly conservedgrowth mediafunction analysisclinical antibioticscentral metabolismcatalytic inhibitorantibiotic productionactive site>- acetylglucosamine6p dehydratase<p><b>(a)</b> Sensitivity of <i>S. coelicolor</i> mutants to GlcNAc. Spores (5 × 10<sup>5</sup> CFU) of <i>S. coelicolor</i> M145 and its mutant derivatives ∆<i>nagB</i>, SMA11, ∆<i>nagB</i>∆<i>nagS</i>, ∆<i>nagB</i>∆<i>nagS</i><sup>C</sup> (∆<i>nagB</i>∆<i>nagS</i> expressing <i>nagS</i>) and ∆<i>nagB</i>∆<i>nagS</i><sup>E</sup> (∆<i>nagB</i>∆<i>nagS</i> with empty plasmid pSET152) were streaked on MM agar plates with 1% mannitol (Mann) and 1% mannitol plus 10 mM GlcNAc (GlcNAc). Strains were cultured for 72 h at 30 °C. Note that <i>nagB</i> mutants can grow in the presence of 10 mM GlcNAc only when SCO4393 (<i>nagS)</i> has been mutated (suppressor SMA11) or deleted. <b>(b)</b> NagS and its role in GlcNAc sensing. Spores of M145 and ∆<i>nagS</i> were plated on MM and R5 with 0, 0.001, 0.01, 0.1, 1, 5, 10, 20, 50, 100, 150, and 200 mM GlcNAc. Note that <i>nagS</i> mutants hardly respond to GlcNAc. <b>(c)</b> Phylogenetic tree of several different types of sugar isomerase (SIS) domain enzymes in bacteria, including NagS, Glucose-6P isomerase (Pgi), Phosphoheptose isomerase (GmhA), <i>N</i>-acetylmuramic acid-6-phosphate etherase (MurQ), Glutamine-fructose-6-phosphate aminotransferase (GlmS), and putative D-galactosamine-6-phosphate deaminase (Agas). The phylogenetic tree was made by MEGA11 (Neighbour-joining method) and built based on the alignment of the amino acid sequences. <b>(d)</b> ITC analysis of NagS with <i>N</i>-acetyl-6-phosphate amino sugar metabolites, namely GlcNAc-6P, ManNAc-6P, and GalNAc-6P. Both GlcNAc-6P and ManNAc-6P are bound well by NagS, while the enzyme did not bind to GalNAc-6P. (<b>e and f</b>) confirmation of NagS products by LC-MS. Extracted ion chromatograms for GlcNAc-6-P (black trace) and its dehydrated product (pink trace) in the enzymatic reaction mixture of GlcNAc-6-P with either active NagS (e) or heat-inactivated NagS <b>(f)</b>. Peaks relating to the product (<i>m</i>/<i>z</i> = 282.0387) are indicated by red arrows. <b>(g)</b> Kinetics of NagS in the presence of 6-PG (0, 0.33, 1 mM). The <i>V</i><sub>0</sub> data used in d were plotted against the substrate concentration, and each assay was performed in triplicate and expressed as a mean ± standard error. The data underlying this Figure can be found in <a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003514#pbio.3003514.s022" target="_blank">S1 Data</a>.</p>2025-11-25T19:00:32ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pbio.3003514.g001https://figshare.com/articles/figure/NagS_is_a_novel_GlcNAc-6P_dehydratase_/30715199CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/307151992025-11-25T19:00:32Z |
| spellingShingle | NagS is a novel GlcNAc-6P dehydratase. Chao Li (145513) Biochemistry Microbiology Ecology Developmental Biology Inorganic Chemistry Biological Sciences not elsewhere classified controls nutrient signaling also revealed 6 highlighted key residues glcnac sensing requires toxicity pathway dependent 6p deacetylase naga novel glcnac 6 key role substrate glcnac novel enzyme work uncovers wall leads unprecedented reaction substrate mycelium structural analogue streptomycetaceae </ streptomyces </ revolves around promiscuous activity nature ’ n </ multicellular lifestyle metabolic checkpoint medicine makers lytic dismantling landmark event hyphal cell highly conserved growth media function analysis clinical antibiotics central metabolism catalytic inhibitor antibiotic production active site >- acetylglucosamine 6p dehydratase |
| status_str | publishedVersion |
| title | NagS is a novel GlcNAc-6P dehydratase. |
| title_full | NagS is a novel GlcNAc-6P dehydratase. |
| title_fullStr | NagS is a novel GlcNAc-6P dehydratase. |
| title_full_unstemmed | NagS is a novel GlcNAc-6P dehydratase. |
| title_short | NagS is a novel GlcNAc-6P dehydratase. |
| title_sort | NagS is a novel GlcNAc-6P dehydratase. |
| topic | Biochemistry Microbiology Ecology Developmental Biology Inorganic Chemistry Biological Sciences not elsewhere classified controls nutrient signaling also revealed 6 highlighted key residues glcnac sensing requires toxicity pathway dependent 6p deacetylase naga novel glcnac 6 key role substrate glcnac novel enzyme work uncovers wall leads unprecedented reaction substrate mycelium structural analogue streptomycetaceae </ streptomyces </ revolves around promiscuous activity nature ’ n </ multicellular lifestyle metabolic checkpoint medicine makers lytic dismantling landmark event hyphal cell highly conserved growth media function analysis clinical antibiotics central metabolism catalytic inhibitor antibiotic production active site >- acetylglucosamine 6p dehydratase |