Bound-state residual dipolar couplings for rapidly exchanging ligands of His-tagged proteins.

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Seidel RD 3rd, Zhuang T, Prestegard JH

Bound-state residual dipolar couplings for rapidly exchanging ligands of His-tagged proteins.

J Am Chem Soc. 2007 Apr 18;129(15):4834-9. Epub 2007 Mar 27.

PubMed ID

17385862 [ View in PubMed

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Abstract

The study of bound-state conformations of ligands interacting with proteins is important to the understanding of protein function and the design of drugs that alter function. Traditionally, transferred nuclear Overhauser effects (trNOEs), measured from NMR spectra of ligands in rapid exchange between bound and free states, have been used in these studies, owing to the inherent heavy weighting of bound-state data in the averaged ligand signals. In principle, residual dipolar couplings (RDCs) provide a useful complement to NOE data in that they provide orientational constraints as opposed to distance constraints, but use in ligand-binding applications has been limited due to the absence of heavy weighting of bound-state data. A widely applicable approach to increasing the weighting of bound-state data in averaged RDCs measured on ligands is presented. The approach rests on association of a His-tagged protein with a nickel-chelate-carrying lipid inserted into the lipid bilayer-like alignment media used in the acquisition of RDCs. The approach is validated through the observation of bound-state RDCs for the disaccharide, lactose, bound to the carbohydrate recognition domain of the mammalian lectin, galectin-3.

DrugBank Data that Cites this Article

Drug Targets

Drug	Target	Kind	Organism	Pharmacological Action	Actions
Lactose	Galectin-3	Protein	Humans	Unknown	Not Available	Details