Kinetic and thermodynamic insights into the inhibitory mechanism of TMG-chitotriomycin on Vibrio campbellii GH20 exo-β-N-acetylglucosaminidase.
In: Carbohydrate research, Jg. 499 (2021), S. 108201
Online
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Zugriff:
We investigated the inhibition kinetics of VhGlcNAcase, a GH20 exo-β-N-acetylglucosaminidase (GlcNAcase) from the marine bacterium Vibrio campbellii (formerly V. harveyi) ATCC BAA-1116, using TMG-chitotriomycin, a natural enzyme inhibitor specific for GH20 GlcNAcases from chitin-processing organisms, with p-nitrophenyl N-acetyl-β-d-glucosaminide (pNP-GlcNAc) as the substrate. TMG-chitotriomycin inhibited VhGlcNAcase with an IC 50 of 3.0 ± 0.7 μM. Using Dixon plots, the inhibition kinetics indicated that TMG-chitotriomycin is a competitive inhibitor, with an inhibition constant K i of 2.2 ± 0.3 μM. Isothermal titration calorimetry experiments provided the thermodynamic parameters for the binding of TMG-chitotriomycin to VhGlcNAcase and revealed that binding was driven by both favorable enthalpy and entropy changes (ΔH° = -2.5 ± 0.1 kcal/mol and -TΔS° = -5.8 ± 0.3 kcal/mol), resulting in a free energy change, ΔG°, of -8.2 ± 0.2 kcal/mol. Dissection of the entropic term showed that a favorable solvation entropy change (-TΔS solv ° = -16 ± 2 kcal/mol) is the main contributor to the entropic term.
(Copyright © 2020 Elsevier Ltd. All rights reserved.)
Titel: |
Kinetic and thermodynamic insights into the inhibitory mechanism of TMG-chitotriomycin on Vibrio campbellii GH20 exo-β-N-acetylglucosaminidase.
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Autor/in / Beteiligte Person: | Morimoto, Y ; Takahashi, S ; Isoda, Y ; Nokami, T ; Fukamizo, T ; Suginta, W ; Ohnuma, T |
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Zeitschrift: | Carbohydrate research, Jg. 499 (2021), S. 108201 |
Veröffentlichung: | Amsterdam : Elsevier ; <i>Original Publication</i>: Amsterdam., 2021 |
Medientyp: | academicJournal |
ISSN: | 1873-426X (electronic) |
DOI: | 10.1016/j.carres.2020.108201 |
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