Rapid Identification of Ligand-Binding Sites by Using an Assignment-Free NMR Approach

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• 作者：Yuya Kodama ; Koh Takeuchi ; Nobuhisa Shimba ; Kohki Ishikawa ; Ei-ichiro Suzuki ; Ichio Shimada ; Hideo Takahashi
• 刊名：Journal of Medicinal Chemistry
• 出版年：2013
• 出版时间：November 27, 2013
• 年：2013
• 卷：56
• 期：22
• 页码：9342-9350
• 全文大小：434K
• 年卷期：v.56,no.22(November 27, 2013)
• ISSN：1520-4804

文摘

In this study, we developed an assignment-free approach for rapid identification of ligand-binding sites in target proteins by using NMR. With a sophisticated cell-free stable isotope-labeling procedure that introduces ¹⁵N- or ¹³C-labels to specific atoms of target proteins, this approach requires only a single series of ligand titrations with labeled targets. Using titration data, ligand-binding sites in the target protein can be identified without time-consuming assignment procedures. We demonstrated the feasibility of this approach by using structurally well-characterized interactions between mitogen-activated protein (MAP) kinase p38伪 and its inhibitor 2-amino-3-benzyloxypyridine. Furthermore, we confirmed the recently proposed fatty acid binding to p38伪 and confirmed the fatty acid-binding site in the MAP kinase insert region.