Applications of NMR and ITC for the Study of the Kinetics of Carbohydrate Binding by AMPK ?-Subunit Carbohydrate-Binding Modules.
Applications of NMR and ITC for the Study of the Kinetics of Carbohydrate Binding by AMPK ?-Subunit Carbohydrate-Binding Modules.
Methods Mol Biol. 2018;1732:87-98
Authors: Gooley
PR, Koay A, Mobbs JI
Abstract
Understanding the kinetics of proteins interacting with their ligands is important for characterizing molecular mechanism. However, it can be difficult to determine the extent and nature of these interactions for weakly formed protein-ligand complexes that have lifetimes of micro- to milliseconds. Nuclear magnetic resonance (NMR) spectroscopy is a powerful solution-based method for the atomic-level analysis of molecular interactions on a wide range of timescales, including micro- to milliseconds. Recently the combination of thermodynamic experiments using isothermal titration calorimetry (ITC) with kinetic measurements using ZZ-exchange and CPMG relaxation dispersion NMR spectroscopy have been used to determine the kinetics of weakly interacting protein systems. This chapter describes the application of ITC and NMR to understand the differences in the kinetics of carbohydrate binding by the ?1- and ?2-carbohydrate-binding modules of AMP-activated protein kinase.
PMID: 29480470 [PubMed - in process]
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