A High Capacity Screen for Membrane-Active Compound(RMI)

描述(申请人提供)：膜蛋白是大多数药物的靶标。其中许多药物深入结合在膜的非极性区域，在那里跨膜螺旋(TM)相互作用是目标蛋白质稳定性和活性的主导因素。我们开发的分析方法可能有助于寻找特定影响TM-TM相互作用的分子，加强或削弱这些关键结构元素的关联。如果能够在筛查中发现这种分子，它们将为发现新的药理制剂提供线索。为了研究这种可能性，我们将使用我们已经研究过并已知通过其TM结构域相互作用的重要蛋白质进行筛选：PDGF受体TM，来自乳头瘤病毒的E5蛋白的TM，PDGFR TM与E5的相互作用，以及来自MHC的Li TM。我们将通过不同靶点之间的交叉比较，通过与Glycophorin A TM的阳性和阴性对照，以及通过对最特异和有效的点击的TM突变分析来测试特异性，其中特定结合部位的修饰将改变亲和力和有效性。

DESCRIPTION (provided by applicant): Membrane proteins are the targets of a majority of all drugs. Many of these drugs bind deeply within the non-polar region of the membrane, where transmembrane helix (TM) interactions are a dominant factor in the stability and activity of target proteins. We have developed assays that might be useful in finding molecules that specifically influence TM-TM interactions, either strengthening or weakening the association of these key elements of structure. If such molecules can be found in screens, they will provide leads for the discovery of new pharmacological agents. To investigate this possibility, we will set up screens using important proteins that we have worked with and already know to interact through their TM domains: the PDGF receptor TM, the TM of the E5 protein from Papilloma virus, the interaction of PDGFR TM with E5, and the li TM from the MHC. We will test specificity by cross comparison among different targets, by positive and negative controls with Glycophorin A TM, and by TM mutational analysis of the most specific and effective hits, where it is expected that modification of a specific binding site will alter affinity and efficacy.