Liposome Overlays For Protein-Ligand Selectivity

描述（由申请人提供）：磷酸肌醇（PIPnS）是细胞通讯的整合信号分子。通过将特异性蛋白质募集到内膜，然后启动信号级联，导致下游效应，如蛋白质分选、肿瘤发生、外吞和内吞作用的调节、细胞形状的控制、细胞迁移和增殖以及凋亡。发现新的PIPn特异性脂质识别蛋白（LRP）在产生用于药物发现的新的潜在靶点方面变得重要。 脂质选择性的生物化学研究已经开始采用蛋白质覆盖脂质印迹，俗称“脂肪蛋白质印迹”，与硝化纤维素固定的PIPns。匡威的方法将采用脂质覆盖凝胶分离的蛋白质。因此，在第一阶段，我们采用了脂质体覆盖系统，用于确定固定在硝酸纤维素膜上的PIPn结合蛋白，使用传统的含PIPn的脂质体（PIPosomesTM）。此外，我们开发了一套新的含有PIPn的聚合脂质体，命名为PolyPIPosomes TM。对于几个PIPn头基，我们还研究了使用PolyPIPosomes TM检测脂质-蛋白质结合相互作用。 在第二阶段，我们建议扩展该方法，以包括所有八个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，其中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β测试产品的合作，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.