Identification and specificity studies of small-molecule ligands for SH3 protein domains
Identification and specificity studies of small-molecule ligands for SH3 protein domains
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DOI：
10.1021/jm049533z
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发表时间：
2004-10-21
影响因子：
7.3
通讯作者：
Booker, GW
中科院分区：
文献类型：
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作者：
Inglis, SR;Stojkoski, C;Booker, GW
Src同源3（SH 3）结构域是小的蛋白质-蛋白质相互作用结构域，其结合富含脯氨酸的序列并介导广泛的细胞信号传导和其他重要的生物学过程。由于失调的信号传导途径形成许多人类疾病的基础，因此SH 3结构域已成为新疗法的有吸引力的靶标。已经设计了针对SH 3结构域的高亲和力配体;然而，这些都是基于肽的，并且以前没有报道过完全非肽SH 3配体的例子。使用小鼠Tec激酶SH 3结构域作为基于结构的配体设计的模型系统，我们已经确定了几个简单的杂环化合物，选择性地结合到Tec SH 3结构域。使用核磁共振化学位移扰动，结构-活性关系，和定点诱变的组合，这些化合物在富含脯氨酸的肽结合位点的结合已被表征。其中最有效的，2-氨基喹啉，与Kd = 125 μ M的结合，并能够竞争与脯氨酸丰富的肽结合。6-取代的-2-氨基喹啉的合成导致配体的亲和力比2-氨基喹啉提高了6倍，并且增强了对Tec SH 3结构域的特异性。因此，2-氨基喹啉可能是潜在的用于开发高亲和力的小分子配体的SH 3结构域。
The Src Homology 3 (SH3) domains are small protein-protein interaction domains that bind proline-rich sequences and mediate a wide range of cell-signaling and other important biological processes. Since deregulated signaling pathways form the basis of many human diseases, the SH3 domains have been attractive targets for novel therapeutics. High-affinity ligands for SH3 domains have been designed; however, these have all been peptide-based and no examples of entirely nonpeptide SH3 ligands have previously been reported. Using the mouse Tec Kinase SH3 domain as a model system for structure-based ligand design, we have identified several simple heterocyclic compounds that selectively bind to the Tec SH3 domain. Using a combination of nuclear magnetic resonance chemical shift perturbation, structure-activity relationships, and site-directed mutagenesis, the binding of these compounds at the proline-rich peptide-binding site has been characterized. The most potent of these, 2-aminoquinoline, bound with K-d = 125 muM and was able to compete for binding with a proline-rich peptide. Synthesis of 6-substitued-2-aminoquinolines resulted in ligands with up to 6-fold improved affinity over 2-aminoquinoline and enhanced specificity for the Tec SH3 domain. Therefore, 2-aminoquinolines may potentially be useful for the development of high affinity small molecule ligands for SH3 domains.