Clean STD-NMR spectrum for improved detection of ligand-protein interactions at low c

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Clean STD-NMR spectrum for improved detection of ligand-protein interactions at low concentration of protein.

Related Articles Clean STD-NMR spectrum for improved detection of ligand-protein interactions at low concentration of protein.

Magn Reson Chem. 2010 Oct 18;

Authors: Xia Y, Zhu Q, Jun KY, Wang J, Gao X

Saturation transfer difference (STD)-NMR has been widely used to screen ligand compound libraries for their binding activities to proteins and to determine the binding epitopes of the ligands. We report herein, a Clean STD-NMR method developed to overcome false positives (artifacts) observed in the STD-NMR spectrum due to the power spillover of RF irradiation. The method achieved higher degree of resonance saturation through digital editing of two STD-NMR spectra to generate a concatenated difference spectrum and three times of sensitivity enhancement for a loose binding complex involving DNA oligonucleotide and an RNA-binding protein, CUGBP-1ab (25.2 kDa). The interesting binding characteristics of the complex dCTGTCT-CUGBP1ab were obtained. The method was applied to a mixture of small ligand and bovine serum albumin protein (BSA, 66.3 kDa), and detected the intermolecular contacts at a BSA concentration as low as 0.1 µM, a working concentration useful for the detection of proteins of low solubility at biologically relevant conditions. Copyright © 2010 John Wiley & Sons, Ltd.

PMID: 20957656 [PubMed - as supplied by publisher]



Source: PubMed