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

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• 作者：Anasztázia Hetényi ; Zsófia Hegedűs ; Roberta Fajka-Boja…
• 关键词：GS ; STD ; Protein mixture ; Antibody ; Protein–ligand interaction ; Gal ; 1
• 刊名：Journal of Biomolecular NMR
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：66
• 期：4
• 页码：227-232
• 全文大小：
• 刊物类别：Physics and Astronomy
• 刊物主题：Biological and Medical Physics, Biophysics; Biochemistry, general; Spectroscopy/Spectrometry;
• 出版者：Springer Netherlands
• ISSN：1573-5001
• 卷排序：66

文摘

Fragment-based drug design has been successfully applied to challenging targets where the detection of the weak protein–ligand interactions is a key element. 1H 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 15N-GS STD spectroscopy has been developed to resolve the problem of protein mixtures and impure proteins. A 15N-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.