2017年4月
NMR line shape analysis of a multi-state ligand binding mechanism in chitosanase
JOURNAL OF BIOMOLECULAR NMR
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- 巻
- 67
- 号
- 4
- 開始ページ
- 309
- 終了ページ
- 319
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1007/s10858-017-0109-6
- 出版者・発行元
- SPRINGER
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.
- リンク情報
- ID情報
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- DOI : 10.1007/s10858-017-0109-6
- ISSN : 0925-2738
- eISSN : 1573-5001
- PubMed ID : 28393280
- Web of Science ID : WOS:000400891000005