Immobilized Proteins In Drug Discovery

基于在流动色谱系统中包含靶受体，跨膜受体（细胞、核或线粒体），已经开发了用于高通量筛选的新方法。在这种方法中，靶蛋白被固定在固体支持物上，并将支持物填充到小柱中。测试化学品通过柱，经过固定的靶标，并且化合物从柱的开始到其结束所花费的时间与靶标和化合物之间的相互作用的强度，即配体-受体复合物的结合亲和力直接相关。使用这种方法，复杂的化学和生物混合物可以在与疾病相关靶标相互作用和不相互作用的化合物之间快速分类。与此同时，与靶标结合的化合物本身在低、中和高亲和力结合剂之间快速分选。因此，该方法快速地提供具有高信息含量的大量数据。我们已经开发了包含烟碱受体的各种亚型的柱来筛选烟草烟雾冷凝物。 一般方法也已扩展到将SIRT 6蛋白固定到开放的管状毛细管表面和磁珠表面上。 SIRT 6-MB用于从化学和植物混合物中提取与SIRT 6结合的小分子。开发并优化了另外的SIRT 6活性测定以鉴定SIRT 6激活剂和SIRT 6抑制剂。使用这种方法，已经从复杂的基质中鉴定出几种新型SIRT 6激活剂。并利用其药理活性建立药效团模型。最近，rhBri 2也被固定在磁珠表面。Bri 2-固定化的蛋白质用于从复杂基质中鉴定相互作用的蛋白质。

A new method for high throughput screening has been developed based upon the inclusion of the target receptors, transmembrane receptors (cellular, nuclear or mitochondrial) in a flowing chromatographic system. In this approach, the target protein is immobilized on a solid support and the support packed into a small column. The test chemicals are passed through the column, over the immobilized target, and the time that it takes for the compounds to pass from the beginning of the column to its end is directly related to the strength of interaction between the target and the compound, i.e. the binding affinity of the ligand-receptor complex. Using this method, complex chemical and biological mixtures can be rapidly sorted between compounds that interact and do not interact with the disease-related target. At the same time, the compounds that bind to the target are themselves rapidly sorted between low, medium and high affinity binders. Thus, the method quickly provides a large amount of data with high information content. We have developed columns containing various subtypes of the nicotinic receptor to screen tobacco smoke condensates. The general approach has also been expanded to the immobilization of SIRT6 protein onto the surface of an open tubular capillary and on the surface of magnetic beads. The SIRT6-MBs were used to extract small molecules that bind to SIRT6 from chemical and botanical mixtures. Additional SIRT6 activity assays were developed and optimized for the identification of SIRT6 activators and SIRT6 inhibitors. Using this approach, several novel SIRT6 activators have been identified from complex matrixes. And the pharmacological activity was used to create a pharmacophore model. More recently, rhBri2 was also immobilized onto the surface of magnetic beads. The Bri2-immobilized proteins was used to identify interacting proteins from a complex matrix.