Document Type
Article
Language
eng
Publication Date
4-2017
Publisher
Springer
Source Publication
Journal of Biomolecular NMR
Source ISSN
0925-2738
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.
Recommended Citation
Shinya, Shoko; Ghinet, Mariana G.; Brzezinski, Ryszard; Furuita, Kyoko; Kojima, Chojiro; Shah, Sneha; Kovrigin, Evgeni; and Fukamizo, Tamo, "NMR Line Shape Analysis of a Multi-state Ligand Binding Mechanism in Chitosanase" (2017). Chemistry Faculty Research and Publications. 583.
https://epublications.marquette.edu/chem_fac/583
Comments
Accepted version. Journal of Biomolecular NMR, Vol. 67, No. 4 (April 2017): 309-319. DOI. © 2017 Springer International Publishing AG. Part of Springer Nature. Used with permission.