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Default NMR line shape analysis of a multi-state ligand binding mechanism in chitosanase.

NMR line shape analysis of a multi-state ligand binding mechanism in chitosanase.

Related Articles NMR line shape analysis of a multi-state ligand binding mechanism in chitosanase.

J Biomol NMR. 2017 Apr 09;:

Authors: Shinya S, Ghinet MG, Brzezinski R, Furuita K, Kojima C, Shah S, Kovrigin EL, Fukamizo T

Abstract
Chitosan interaction with chitosanase was examined through analysis of spectral line shapes in the NMR HSQC titration experiments. We established that the substrate, chitosan hexamer, binds to the enzyme through the three-state induced-fit mechanism with fast formation of the encounter complex followed by slow isomerization of the bound-state into the final conformation. Mapping of the chemical shift perturbations in two sequential steps of the mechanism highlighted involvement of the substrate-binding subsites and the hinge region in the binding reaction. Equilibrium parameters of the three-state model agreed with the overall thermodynamic dissociation constant determined by ITC. This study presented the first kinetic evidence of the induced-fit mechanism in the glycoside hydrolases.


PMID: 28393280 [PubMed - as supplied by publisher]



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