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.
Ligand Binding Properties of the Lentil Lipid TransferProtein: Molecular Insight into the Possible Mechanism of Lipid Uptake
Ligand Binding Properties of the Lentil Lipid TransferProtein: Molecular Insight into the Possible Mechanism of Lipid Uptake
http://pubs.acs.org/appl/literatum/publisher/achs/journals/content/bichaw/0/bichaw.ahead-of-print/acs.biochem.6b01079/20170316/images/medium/bi-2016-010796_0006.gif
Biochemistry
DOI: 10.1021/acs.biochem.6b01079
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Predicting the redox state and secondary structure of cysteine residues using multi-dimensional classification analysis of NMR chemical shifts
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Abstract
A tool for predicting the redox state and secondary structure of cysteine residues using multi-dimensional analyses of different combinations of nuclear magnetic resonance (NMR) chemical shifts has been developed. A data set of cysteine \({}^{13}{\text{C}}^{'}\) , 13Cα, 13Cβ, 1Hα, 1HN, and 15NH chemical shifts was...
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Abstract
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[NMR paper] Mass spectrometry and NMR analysis of ligand binding by human liver fatty acid binding protein.
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Abstract
Human liver fatty acid binding protein (hL-FABP) is the most abundant cytosolic protein in the liver. This protein plays important roles associated to partitioning of fatty acids (FAs) to specific...
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Abstract Biological function of proteins relies on conformational transitions and binding of specific ligands. Proteinâ??ligand interactions are thermodynamically and kinetically coupled to conformational changes in protein structures as conceptualized by the models of pre-existing equilibria and induced fit. NMR spectroscopy is particularly sensitive to complex ligand-binding modesâ??NMR line-shape analysis can provide for thermodynamic and kinetic constants of ligand-binding equilibria with the site-specific resolution. However,...
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We have recently proposed and demonstrated an approach that enables the acquisition of 2D nuclear magnetic resonance (NMR) spectra within a single scan. The approach is based on spatially encoding the spins' evolution along the indirect domain with the aid of a magnetic field gradient, and subsequently decoding this information numerous times over...
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Octopine dehydrogenase (OcDH) from the adductor muscle of the great scallop, Pecten maximus, catalyzes the NADH dependent, reductive condensation of L-arginine and pyruvate to...
NMR line shape analysis of a multi-state ligand binding mechanism in chitosanase
Linear Mode
