EphA2 receptor plays a critical and debatable function in cancer and is considered a target in drug discovery. Lately, there has been a growing interest in its cytosolic C-terminal SAM domain (EphA2-SAM) as it engages protein modulators of receptor endocytosis and stability. Interestingly, EphA2-SAM binds the SAM domain from the lipid phosphatase Ship2 (Ship2-SAM) mainly producing pro-oncogenic outcomes. In an attempt to discover novel inhibitors of the EphA2-SAM/Ship2-SAM complex with possible anticancer properties, we focused on the central region of Ship2-SAM (known as Mid-Loop interface) responsible for its binding to EphA2-SAM. Starting from the amino acid sequence of the Mid-Loop interface virtual peptide libraries were built through ad hoc inserted mutations with either l- or d- amino acids and screened against EphA2-SAM by docking techniques. A few virtual hits were synthesized and experimentally tested by a variety of direct and competition-type interaction assays relying on NMR (Nuclear Magnetic Resonance), SPR (Surface Plasmon Resonance), MST (Microscale Thermophoresis) techniques. These studies guided the discovery of an original EphA2-SAM ligand antagonist of its interaction with Ship2-SAM.

中文翻译：

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EphA2-SAM肽配体的设计和分析：多学科筛选方法。

EphA2受体在癌症中起着至关重要的作用，被认为是药物开发的目标。最近，人们越来越关注其胞质C端SAM结构域（EphA2-SAM），因为它参与了受体内吞作用和稳定性的蛋白质调节剂。有趣的是，EphA2-SAM与脂质磷酸酶Ship2（Ship2-SAM）的SAM结构域结合，主要产生促癌作用。为了尝试发现具有可能的抗癌特性的EphA2-SAM / Ship2-SAM复合物的新型抑制剂，我们集中于Ship2-SAM的中央区域（称为Mid-Loop接口）负责与EphA2-SAM的结合。从Mid-Loop接口的氨基酸序列开始，通过具有l或d-氨基酸的ad hoc插入突变构建虚拟肽库，并通过对接技术针对EphA2-SAM进行筛选。依靠NMR（核磁共振），SPR（表面等离振子共振），MST（微尺度热泳）技术，通过各种直接和竞争型相互作用测定法合成了一些虚拟命中物并进行了实验测试。这些研究指导了其与Ship2-SAM相互作用的原始EphA2-SAM配体拮抗剂的发现。