Molecular Recognition in Biomimetic Receptors

描述（由申请人提供）：本申请的主要目的是开发能够识别蛋白质外表面并破坏临床上重要的蛋白质-蛋白质相互作用的合成试剂。我们将利用每个蛋白质表面上带电、疏水和亲水基团的独特分布来识别选择性结合并可能破坏生物功能的人工受体。我们的方法的一个中心特征将是设计合成试剂，其包含大的（>400埃2）和功能化的表面积以识别靶蛋白的互补表面。在前一个项目期间，我们已经表明，这种策略可以产生对特定蛋白质靶点表现出高亲和力（nM-pM）的蛋白质结合剂，在一系列不同蛋白质中具有选择性，在高盐浓度条件下有效，并且可以破坏体内蛋白质-蛋白质相互作用。在这些主要目标范围内，我们将有以下具体目标： 具体目标1-开发与异常细胞增殖途径中涉及的生长因子蛋白（包括PDGF、VEGF、EGF和FGF）具有高亲和力和选择性结合的合成受体。 具体目标2-开发与细胞色素c表面结合并破坏其与Apaf-1相互作用以形成线粒体的合成试剂。我们将靶向细胞色素c的外表面，该表面参与其与蛋白质伴侣的相互作用。 具体目标3-开发选择性结合磷酸化蛋白质表面的合成配体，并通过荧光发射变化发出信号。我们将使用环状三联苯酰胺支架开发靶向活化信号蛋白（如Bcr-Abl，STAT 3和Src）表面磷酸酪氨酸基团的结合剂。具体目标4-开发可用于检测具有不同表面性质的蛋白质并优化其他靶标的结合选择性的荧光结合剂阵列。我们将开发荧光蛋白结合剂的溶液和固相阵列，这些荧光蛋白结合剂在具有不同表面识别特性的蛋白质存在下显示出独特的淬灭模式。

DESCRIPTION (provided by applicant): The primary aims of this application will be to develop synthetic agents that can recognize the exterior surface of proteins and disrupt clinically important protein-protein interactions. We will exploit the unique distribution of charged, hydrophobic and hydrophilic groups on the surface of every protein to identify artificial receptors that bind selectively and potentially disrupt biological function. A central feature of our approach will be the design of synthetic agents that contain a large (>400 Angstrom 2) and functionalized surface area to recognize the complementary surface of the target protein. In the previous project period we have shown that this strategy can lead to protein binding agents that show high affinity (nM-pM) for specific protein targets, are selective among a range of different proteins, are effective in conditions of high salt concentration and can disrupt protein-protein interactions in vivo. Within these primary goals we will have the following specific aims: Specific Aim 1- To develop synthetic receptors that bind with high affinity and selectivity to growth factor proteins (including PDGF, VEGF, EGF and FGF) involved in aberrant cell proliferation pathways. Specific Aim 2- To develop synthetic agents that bind to the surface of cytochrome c and disrupt its interaction with Apaf-1 to form the apoptosome. We will target the exterior surface of cytochrome c that is involved in its interactions with protein partners. Specific Aim 3- To develop synthetic ligands that bind selectively to the surface of phosphorylated proteins and signal their presence through fluorescence emission changes. We will develop binding agents that target the phosphotyrosine groups on the surface of activated signaling proteins, such as Bcr-Abl, STAT3 and Src using a cyclic tris-biphenylamide scaffold. Specific Aim 4- To develop arrays of fluorescent binding agents that can be used to detect proteins with different surface properties and to optimize binding selectivities for other targets. We will develop solution and solid phase arrays of fluorescent protein binding agents that show distinctive patterns of quenching in the presence of proteins with different surface recognition characteristics.