The focal adhesion kinase (FAK) and the proline‐rich tyrosine kinase 2‐beta (PYK2) are implicated in cancer progression and metastasis and represent promising biomarkers and targets for cancer therapy. FAK and PYK2 are recruited to focal adhesions (FAs) via interactions between their FA targeting (FAT) domains and conserved segments (LD motifs) on the proteins Paxillin, Leupaxin, and Hic‐5. A promising new approach for the inhibition of FAK and PYK2 targets interactions of the FAK domains with proteins that promote localization at FAs. Advances toward this goal include the development of surface plasmon resonance, heteronuclear single quantum coherence nuclear magnetic resonance (HSQC‐NMR) and fluorescence polarization assays for the identification of fragments or compounds interfering with the FAK‐Paxillin interaction. We have recently validated this strategy, showing that Paxillin mimicking polypeptides with 2 to 3 LD motifs displace FAK from FAs and block kinase‐dependent and independent functions of FAK, including downstream integrin signaling and FA localization of the protein p130Cas. In the present work we study by all‐atom molecular dynamics simulations the recognition of peptides with the Paxillin and Leupaxin LD motifs by the FAK‐FAT and PYK2‐FAT domains. Our simulations and free‐energy analysis interpret experimental data on binding of Paxillin and Leupaxin LD motifs at FAK‐FAT and PYK2‐FAT binding sites, and assess the roles of consensus LD regions and flanking residues. Our results can assist in the design of effective inhibitory peptides of the FAK‐FAT: Paxillin and PYK2‐FAT:Leupaxin complexes and the construction of pharmacophore models for the discovery of potential small‐molecule inhibitors of the FAK‐FAT and PYK2‐FAT focal adhesion based functions.

中文翻译：

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通过粘着斑激酶和富含脯氨酸的酪氨酸激酶 2-β 的粘着斑靶向域识别 LD 基序：来自分子动力学模拟的见解。


粘着斑激酶 (FAK) 和富含脯氨酸的酪氨酸激酶 2-β (PYK2) 与癌症进展和转移有关，是癌症治疗的有前途的生物标志物和靶点。 FAK 和 PYK2 通过其 FA 靶向 (FAT) 结构域与 Paxillin、Leupaxin 和 Hic-5 蛋白上的保守片段（LD 基序）之间的相互作用而被招募到粘着斑 (FA)。一种有前景的抑制 FAK 和 PYK2 的新方法以 FAK 结构域与促进 FA 定位的蛋白质相互作用为目标。实现这一目标的进展包括表面等离子体共振、异核单量子相干核磁共振 (HSQC-NMR) 和荧光偏振测定的发展，用于识别干扰 FAK-Paxillin 相互作用的片段或化合物。我们最近验证了这一策略，表明具有 2 至 3 个 LD 基序的桩蛋白模拟多肽可取代 FA 中的 FAK，并阻断 FAK 的激酶依赖性和独立功能，包括下游整联蛋白信号传导和蛋白质 p130Cas 的 FA 定位。在目前的工作中，我们通过全原子分子动力学模拟研究 FAK-FAT 和 PYK2-FAT 结构域对具有 Paxillin 和 Leupaxin LD 基序的肽的识别。我们的模拟和自由能分析解释了 Paxillin 和 Leupaxin LD 基序在 FAK-FAT 和 PYK2-FAT 结合位点上结合的实验数据，并评估了共有 LD 区域和侧翼残基的作用。我们的结果可以帮助设计 FAK-FAT: Paxillin 和 PYK2-FAT:Leupaxin 复合物的有效抑制肽，以及构建药效团模型，以发现 FAK-FAT 和 PYK2-FAT 焦点的潜在小分子抑制剂。基于粘附的功能。