Size-exclusion chromatography analysis of purified NagS.
<p>Analytical gel-filtration chromatography was performed on a Superdex 200 10/300 GL column (Cytiva) equilibrated with buffer containing 20 mM HEPES pH 7.5, 300 mM NaCl, 5% glycerol, and 1 mM DTT. NagS was analyzed at two protein concentrations (35 μM and 70 μM). Elution volumes (V<sub>...
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2025
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| _version_ | 1849927625880895488 |
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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 | 2025-11-25T19:00:09Z |
| dc.identifier.none.fl_str_mv | 10.1371/journal.pbio.3003514.s005 |
| dc.relation.none.fl_str_mv | https://figshare.com/articles/figure/Size-exclusion_chromatography_analysis_of_purified_NagS_/30715144 |
| 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 | Size-exclusion chromatography analysis of purified NagS. |
| dc.type.none.fl_str_mv | Image Figure info:eu-repo/semantics/publishedVersion image |
| description | <p>Analytical gel-filtration chromatography was performed on a Superdex 200 10/300 GL column (Cytiva) equilibrated with buffer containing 20 mM HEPES pH 7.5, 300 mM NaCl, 5% glycerol, and 1 mM DTT. NagS was analyzed at two protein concentrations (35 μM and 70 μM). Elution volumes (V<sub>e</sub>) (<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003514#pbio.3003514.s021" target="_blank">S7 Table</a>) were compared to a molecular weight (Mw) calibration curve above generated using a Protein Standard Mix under identical buffer conditions. The calibration curve followed the equation V<sub>e</sub> = –1.525ln(Mw) + 20.659 (R² = 0.964). Note: NagS eluted at 14.45 ml, which corresponds to an estimated molecular weight of ~60.04 kDa according to the calibration curve equation. Given that the calculated molecular weight of a NagS-His₆ monomer is 28.29 kDa, these results indicate that NagS exists as a dimer in the tested buffer. 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> <p>(TIF)</p> |
| eu_rights_str_mv | openAccess |
| id | Manara_319440e7046ade2d183e05f4b12fea26 |
| identifier_str_mv | 10.1371/journal.pbio.3003514.s005 |
| network_acronym_str | Manara |
| network_name_str | ManaraRepo |
| oai_identifier_str | oai:figshare.com:article/30715144 |
| publishDate | 2025 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| rights_invalid_str_mv | CC BY 4.0 |
| spelling | Size-exclusion chromatography analysis of purified NagS.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>Analytical gel-filtration chromatography was performed on a Superdex 200 10/300 GL column (Cytiva) equilibrated with buffer containing 20 mM HEPES pH 7.5, 300 mM NaCl, 5% glycerol, and 1 mM DTT. NagS was analyzed at two protein concentrations (35 μM and 70 μM). Elution volumes (V<sub>e</sub>) (<a href="http://www.plosbiology.org/article/info:doi/10.1371/journal.pbio.3003514#pbio.3003514.s021" target="_blank">S7 Table</a>) were compared to a molecular weight (Mw) calibration curve above generated using a Protein Standard Mix under identical buffer conditions. The calibration curve followed the equation V<sub>e</sub> = –1.525ln(Mw) + 20.659 (R² = 0.964). Note: NagS eluted at 14.45 ml, which corresponds to an estimated molecular weight of ~60.04 kDa according to the calibration curve equation. Given that the calculated molecular weight of a NagS-His₆ monomer is 28.29 kDa, these results indicate that NagS exists as a dimer in the tested buffer. 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> <p>(TIF)</p>2025-11-25T19:00:09ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pbio.3003514.s005https://figshare.com/articles/figure/Size-exclusion_chromatography_analysis_of_purified_NagS_/30715144CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/307151442025-11-25T19:00:09Z |
| spellingShingle | Size-exclusion chromatography analysis of purified NagS. 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 | Size-exclusion chromatography analysis of purified NagS. |
| title_full | Size-exclusion chromatography analysis of purified NagS. |
| title_fullStr | Size-exclusion chromatography analysis of purified NagS. |
| title_full_unstemmed | Size-exclusion chromatography analysis of purified NagS. |
| title_short | Size-exclusion chromatography analysis of purified NagS. |
| title_sort | Size-exclusion chromatography analysis of purified NagS. |
| 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 |