Liposome Overlays For Protein-Ligand Selectivity

描述（由申请人提供）：磷酸肌苷（PIPnS）是细胞通讯不可或缺的信号分子。通过将特定的蛋白质招募到内膜，信号级联被启动，导致下游效应，如蛋白质分选、肿瘤发生、外胞和内吞作用的调节、细胞形状的控制、细胞迁移和增殖以及细胞凋亡。发现新的PIPn特异性脂质识别蛋白（lrp）在产生新的潜在药物发现靶点方面变得非常重要。脂质选择性的生化研究已经开始使用蛋白质覆盖脂质印迹，通俗地称为“Fat western”，与硝化纤维素固定的PIPns。相反的方法将使用脂质覆盖层与凝胶分离的蛋白质。因此，在I期研究中，我们采用脂质体覆盖系统，使用传统的含PIPn脂质体（PIPosomesTM）来鉴定固定在硝化纤维素膜上的PIPn结合蛋白。此外，我们还开发了一套新型的含pipn的聚合脂质体，命名为PolyPIPosomes TM。对于几个PIPn头组，我们还研究了使用polyypiposomes TM检测脂质-蛋白结合相互作用。在II期，我们建议将方法扩展到包括所有8种PIPn支架并进行稳定性研究。此外，我们将进一步探索PolyPIPosomes TM的应用，常规脂质体先前已用于基础研究应用和高通量筛选。最后，我们将开发PolyPIPosomes TM和PIPosomes TM研究产品，并建立合作进行beta测试产品，以促进其推向市场。

DESCRIPTION (provided by applicant): Phosphoinositides (PIPnS) are integral signaling molecules for cellular communication. By recruiting specific proteins to the inner membrane, signal cascades are then initiated leading to downstream effects such as protein sorting, tumorigenesis, regulation of exo-and endocytosis, control of cell shape, cell migration and proliferation, and apoptosis. Discovering new PIPn specific lipid recognition proteins (LRPs) has become important in yielding new potential targets for drug discovery. Biochemical investigations on lipid selectivity have begun to employ protein overlay lipid blots, colloquially termed "Fat Westerns", with nitrocellulose-immobilized PIPns. The converse approach would employ a lipid overlay with gel-separated proteins. Thus, in Phase I, we employed a liposome overlay system for identifying PIPn binding proteins immobilized on a nitrocellulose membrane using traditional PIPn-containing liposomes (PIPosomesTM). In addition, we developed a set of novel PIPn-containing polymerized liposomes named PolyPIPosomes TM. For several of the PIPn headgroups, we also investigated the use of PolyPIPosomes TM for detecting lipid-protein binding interactions. In Phase II, we propose to extend the methodology to include all eight PIPn scaffolds and perform stability studies. In addition we will further explore applications for PolyPIPosomes TM where regular liposomes have previously been used in basic research applications and in high throughput screening. Finally we will develop PolyPIPosomes TM and PIPosomes TM research products and establish collaborations for beta-testing products to facilitate their introduction to the market.