Target-specific NMR detection of protein-ligand interactions with antibody-relayed (15)N-group selective STD.
 

Target-specific NMR detection of protein-ligand interactions with antibody-relayed (15)N-group selective STD.

Target-specific NMR detection of protein-ligand interactions with antibody-relayed (15)N-group selective STD.

J Biomol NMR. 2016 Nov 24;

Authors: Hetényi A, Heged?s Z, Fajka-Boja R, Monostori É, Kövér KE, Martinek TA

Abstract
Fragment-based drug design has been successfully applied to challenging targets where the detection of the weak protein-ligand interactions is a key element. (1)H saturation transfer difference (STD) NMR spectroscopy is a powerful technique for this work but it requires pure homogeneous proteins as targets. Monoclonal antibody (mAb)-relayed (15)N-GS STD spectroscopy has been developed to resolve the problem of protein mixtures and impure proteins. A (15)N-labelled target-specific mAb is selectively irradiated and the saturation is relayed through the target to the ligand. Tests on the anti-Gal-1 mAb/Gal-1/lactose system showed that the approach is experimentally feasible in a reasonable time frame. This method allows detection and identification of binding molecules directly from a protein mixture in a multicomponent system.


PMID: 27885546 [PubMed - as supplied by publisher]



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