Deriving ligand orientation on weak protein-ligand complexes by DEEP-STD NMR in the absence of protein chemical shift assignment

Nepravishta, Ridvan, Walpole, Samuel, Tailford, Louise, Juge, Nathalie and Angulo, Jesus (2019) Deriving ligand orientation on weak protein-ligand complexes by DEEP-STD NMR in the absence of protein chemical shift assignment. ChemBioChem, 20 (3). pp. 340-344. ISSN 1439-4227

[img]
Preview
PDF (Accepted manuscript) - Submitted Version
Available under License Creative Commons Attribution Non-commercial.

Download (1MB) | Preview

Abstract

Differential epitope mapping saturation transfer difference (DEEP-STD) NMR spectroscopy is a recently developed powerful approach for elucidating the structure and pharmacophore of weak protein–ligand interactions, as it reports key information on the orientation of the ligand and the architecture of the binding pocket. The method relies on selective saturation of protein residues in the binding site and the generation of a differential epitope map by observing the ligand, which depicts the nature of the protein residues making contact with the ligand in the bound state. Selective saturation requires knowledge of the chemical-shift assignment of the protein residues, which can be obtained either experimentally by NMR spectroscopy or predicted from 3D structures. Herein, we propose a simple experimental procedure to expand the DEEP-STD NMR methodology to protein–ligand cases in which the spectral assignment of the protein is not available. This is achieved by experimentally identifying the chemical shifts of the residues present in binding hot-spots on the surface of the receptor protein by using 2D NMR experiments combined with a paramagnetic probe.

Item Type: Article
Uncontrolled Keywords: deep-std,nmr spectroscopy,tempol,mixed molecular dynamics,biochemistry,molecular medicine,molecular biology,organic chemistry ,/dk/atira/pure/subjectarea/asjc/1300/1303
Faculty \ School: Faculty of Science > School of Pharmacy
Related URLs:
Depositing User: LivePure Connector
Date Deposited: 01 Nov 2018 09:30
Last Modified: 29 Jul 2020 23:44
URI: https://ueaeprints.uea.ac.uk/id/eprint/68729
DOI: 10.1002/cbic.201800568

Actions (login required)

View Item View Item