Protein-ligand binding affinity determination by the waterLOGSY method: An optimised approach considering ligand rebinding

Renjie Huang; Arnaud Bonnichon; Timothy D W Claridge; Ivanhoe K H Leung

doi:10.1038/srep43727

Protein-ligand binding affinity determination by the waterLOGSY method: An optimised approach considering ligand rebinding

Sci Rep. 2017 Mar 3:7:43727. doi: 10.1038/srep43727.

Authors

Renjie Huang¹, Arnaud Bonnichon^{2

3}, Timothy D W Claridge², Ivanhoe K H Leung¹

Affiliations

¹ School of Chemical Sciences, The University of Auckland, Private Bag 92019, Victoria Street West, Auckland 1142, New Zealand.
² Department of Chemistry, University of Oxford, Chemistry Research Laboratory, 12 Mansfield Road, Oxford OX1 3TA, United Kingdom.
³ Université D'Auvergne, 49 Boulevard François-Mitterrand, CS 60032, Clermont Ferrand 63001, Cedex 1, France.

Abstract

WaterLOGSY is a popular ligand-observed NMR technique to screen for protein-ligand interactions, yet when applied to measure dissociation constants (K_D) through ligand titration, the results were found to be strongly dependent on sample conditions. Herein, we show that accurate K_Ds can be obtained by waterLOGSY with optimised experimental setup.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Caffeine / chemistry
Humans
Kinetics
Ligands*
Nuclear Magnetic Resonance, Biomolecular / methods*
Protein Binding
Proteins / chemistry*
Proteins / metabolism
Serum Albumin / chemistry
Serum Albumin / metabolism
Tryptophan / chemistry

Substances

Ligands
Proteins
Serum Albumin
Caffeine
Tryptophan