Function determination and kinetics of NagS.

(a) 1H 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</...

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主要作者:	Chao Li (145513) (author)
其他作者:	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)
出版:	2025
主題:	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
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總結:	<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>

Function determination and kinetics of NagS.

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