Function determination and kinetics of NagS.

<p>(<b>a</b>) <sup>1</sup>H NMR spectrum of the enzymatic reaction mixture of GlcNAc-6P with either the active NagS (top) or the heated-inactivated one (bottom). The associated NMR peaks of the reaction product are highlighted in red circles and arrows. (<b>b</...

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গ্রন্থ-পঞ্জীর বিবরন
প্রধান লেখক: Chao Li (145513) (author)
অন্যান্য লেখক: Mia Urem (22683580) (author), Ioli Kotsogianni (9930187) (author), Josephine Lau (20442343) (author), Chao Du (288036) (author), Somayah S. Elsayed (9192571) (author), Nathaniel I. Martin (847378) (author), Iain W. McNae (840777) (author), Patrick Voskamp (2311429) (author), Christoph Mayer (57204) (author), Sébastien Rigali (724082) (author), Navraj Pannu (10163763) (author), Jan Pieter Abrahams (1429531) (author), Lennart Schada von Borzyskowski (22683583) (author), Gilles P. van Wezel (7838948) (author)
প্রকাশিত: 2025
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_version_ 1849927625874604032
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 2025-11-25T19:00:11Z
dc.identifier.none.fl_str_mv 10.1371/journal.pbio.3003514.s007
dc.relation.none.fl_str_mv https://figshare.com/articles/figure/Function_determination_and_kinetics_of_NagS_/30715150
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 Function determination and kinetics of NagS.
dc.type.none.fl_str_mv Image
Figure
info:eu-repo/semantics/publishedVersion
image
description <p>(<b>a</b>) <sup>1</sup>H NMR spectrum of the enzymatic reaction mixture of GlcNAc-6P with either the active NagS (top) or the heated-inactivated one (bottom). The associated NMR peaks of the reaction product are highlighted in red circles and arrows. (<b>b</b>) HRESIMS spectrum of the NagS product, compound <b>1</b>: <i>m</i>/<i>z</i> 282.0387 [M-H]<sup>−</sup> (calculated for C<sub>8</sub>H<sub>13</sub>NO<sub>8</sub>P, 282.0384) <b>(c)</b> <sup>1</sup>H NMR spectrum of the enzymatic reaction mixture of ManNAc-6P with either the active NagS (top) or the heat-inactivated one (bottom). The associated NMR peaks of the reaction product are highlighted in red circles. (<b>d</b>) Reactions catalyzed by NagS. NagS dehydrates both GlcNAc-6P and ManNAc-6P to produce compound <b>1</b>. Michaelis-Menten curves were fitted, and selected curves are shown for NagS with the substrates GlcNAc-6P (<b>e</b>) with <i>K</i><sub>m</sub> value of 0.45 ± 0.03 mM and <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 5.48 × 10<sup>4</sup> M<sup>−1</sup>·s<sup>−1</sup>, and ManNAc-6P (<b>f</b>) with <i>K</i><sub>m</sub> value of 0.68 ± 0.05 mM and <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 1.32 × 10<sup>3</sup> M<sup>−1</sup>·s<sup>−1</sup>. In <b>e</b> and <b>f</b>, the <i>V</i><sub>0</sub> data 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> <p>(TIF)</p>
eu_rights_str_mv openAccess
id Manara_1980112c7480c6eee884a29bac6cd352
identifier_str_mv 10.1371/journal.pbio.3003514.s007
network_acronym_str Manara
network_name_str ManaraRepo
oai_identifier_str oai:figshare.com:article/30715150
publishDate 2025
repository.mail.fl_str_mv
repository.name.fl_str_mv
repository_id_str
rights_invalid_str_mv CC BY 4.0
spelling Function determination and kinetics of 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>(<b>a</b>) <sup>1</sup>H NMR spectrum of the enzymatic reaction mixture of GlcNAc-6P with either the active NagS (top) or the heated-inactivated one (bottom). The associated NMR peaks of the reaction product are highlighted in red circles and arrows. (<b>b</b>) HRESIMS spectrum of the NagS product, compound <b>1</b>: <i>m</i>/<i>z</i> 282.0387 [M-H]<sup>−</sup> (calculated for C<sub>8</sub>H<sub>13</sub>NO<sub>8</sub>P, 282.0384) <b>(c)</b> <sup>1</sup>H NMR spectrum of the enzymatic reaction mixture of ManNAc-6P with either the active NagS (top) or the heat-inactivated one (bottom). The associated NMR peaks of the reaction product are highlighted in red circles. (<b>d</b>) Reactions catalyzed by NagS. NagS dehydrates both GlcNAc-6P and ManNAc-6P to produce compound <b>1</b>. Michaelis-Menten curves were fitted, and selected curves are shown for NagS with the substrates GlcNAc-6P (<b>e</b>) with <i>K</i><sub>m</sub> value of 0.45 ± 0.03 mM and <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 5.48 × 10<sup>4</sup> M<sup>−1</sup>·s<sup>−1</sup>, and ManNAc-6P (<b>f</b>) with <i>K</i><sub>m</sub> value of 0.68 ± 0.05 mM and <i>k</i><sub>cat</sub>/<i>K</i><sub>m</sub> value of 1.32 × 10<sup>3</sup> M<sup>−1</sup>·s<sup>−1</sup>. In <b>e</b> and <b>f</b>, the <i>V</i><sub>0</sub> data 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> <p>(TIF)</p>2025-11-25T19:00:11ZImageFigureinfo:eu-repo/semantics/publishedVersionimage10.1371/journal.pbio.3003514.s007https://figshare.com/articles/figure/Function_determination_and_kinetics_of_NagS_/30715150CC BY 4.0info:eu-repo/semantics/openAccessoai:figshare.com:article/307151502025-11-25T19:00:11Z
spellingShingle Function determination and kinetics of 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 Function determination and kinetics of NagS.
title_full Function determination and kinetics of NagS.
title_fullStr Function determination and kinetics of NagS.
title_full_unstemmed Function determination and kinetics of NagS.
title_short Function determination and kinetics of NagS.
title_sort Function determination and kinetics of 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